Pharmaceuticals 07 00545

Pharmaceuticals 2014, 7, 545-594; doi:10.

3390/ph7050545
OPEN ACCESS

pharmaceuticals
ISSN 1424-8247
www.mdpi.com/journal/pharmaceuticals
Review

Human Antimicrobial Peptides and Proteins

Guangshun Wang
Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical

Center, 986495 Nebraska Medical Center, Omaha, NE 68198-6495, USA; E-Mail: [email protected];
Tel.: +402-559-4176; Fax: +402-559-4077.
Received: 17 January 2014; in revised form: 15 April 2014 / Accepted: 29 April 2014 /
Published: 13 May 2014
Abstract: As the key components of innate immunity, human host defense antimicrobial
peptides and proteins (AMPs) play a critical role in warding off invading microbial
pathogens. In addition, AMPs can possess other biological functions such as apoptosis,
wound healing, and immune modulation. This article provides an overview on the
identification, activity, 3D structure, and mechanism of action of human AMPs selected
from the antimicrobial peptide database. Over 100 such peptides have been identified from
a variety of tissues and epithelial surfaces, including skin, eyes, ears, mouths, gut, immune,
nervous and urinary systems. These peptides vary from 10 to 150 amino acids with a net
charge between −3 and +20 and a hydrophobic content below 60%. The sequence diversity
enables human AMPs to adopt various 3D structures and to attack pathogens by different
mechanisms. While α-defensin HD-6 can self-assemble on the bacterial surface into
nanonets to entangle bacteria, both HNP-1 and β-defensin hBD-3 are able to block cell
wall biosynthesis by binding to lipid II. Lysozyme is well-characterized to cleave bacterial
cell wall polysaccharides but can also kill bacteria by a non-catalytic mechanism. The two
hydrophobic domains in the long amphipathic α-helix of human cathelicidin LL-37 lays the
basis for binding and disrupting the curved anionic bacterial membrane surfaces by
forming pores or via the carpet model. Furthermore, dermcidin may serve as ion channel
by forming a long helix-bundle structure. In addition, the C-type lectin RegIIIα can initially
recognize bacterial peptidoglycans followed by pore formation in the membrane. Finally,
histatin 5 and GAPDH(2-32) can enter microbial cells to exert their effects. It appears that
granulysin enters cells and kills intracellular pathogens with the aid of pore-forming
perforin. This arsenal of human defense proteins not only keeps us healthy but also inspires
the development of a new generation of personalized medicine to combat drug-resistant
superbugs, fungi, viruses, parasites, or cancer. Alternatively, multiple factors (e.g., albumin,
Pharmaceuticals 2014, 7 546
arginine, butyrate, calcium, cyclic AMP, isoleucine, short-chain fatty acids, UV B light,
vitamin D, and zinc) are able to induce the expression of antimicrobial peptides, opening
new avenues to the development of anti-infectious drugs.
Keywords: antimicrobial chemokines; antimicrobial neuropeptides; antimicrobial proteins;

cathelicidin LL-37; defensins; dermcidin; hepcidins; histatins; RNases
1. Introduction
Host defense antimicrobial peptides (AMPs) are key components of the innate immune system
shared by both invertebrates and vertebrates. Invertebrates such as insects and crustaceans do not have
adaptive immune systems and innate defense systems serve as the only protective mechanism. It is
now appreciated that innate immune systems also play an indispensable role in vertebrates by directly
killing invading microbes in the early stage. Later, vertebrate AMPs can also help augment the
adaptive immune system to further handle infections. Thus, host defense peptides may have dual role:
rapid microbial killing and subsequent immune modulation [1–6].
The universality of AMPs is evidenced by the identification of these molecules in a variety of
organisms. Over 2,300 such peptides have been isolated and characterized according to the online
updated Antimicrobial Peptide Database (APD) [7,8]. As of April 2014, there are 233 AMPs from
bacteria (i.e., bacteriocins), six from protozoa, 12 from fungi, 306 from plants, and 1,801 host defense
peptides from animals. AMPs are usually gene-coded and can be constitutively expressed or induced
to fend off invading pathogens. In addition, some bacteria use a second mechanism to assemble
peptide antibiotics by using a multiple-enzyme system, leading to distinct chemical modifications not
observed in gene-coded cases. Most of the AMPs are cationic and short with less than 50 amino acids.
Such features are ideal to target the negatively charged surface of bacteria [1–6].
AMPs also protect humans from microbial infection. They have been identified in a variety of
exposed tissues or surfaces such as skin, eyes, ears, mouth, airways, lung, intestines, and the urinary
tract. While human cathelicidin LL-37 is detected in the skin of new born infants [9], human
beta-defensin 2 (hBD-2) is frequently expressed in older individuals. Human S100 proteins, hBD-2,
human beta-defensin 3 (hBD-3), and cathelicidin are significantly higher in fetal keratinocytes than in
postnatal skin cells [10]. In addition, psoriasin (S100A7), RNase 7, and hBD-3 are differentially
expressed in healthy human skin [11]. When the skin barrier is broken, psoriasin is up-regulated [12].
Lysozyme and lactoferrin have been found in human tears [13]. In addition, β-defensins are expressed
in human middle ear epithelial cells [14]. Drosomycin-like defensin (DLD) [15] is produced in human
oral epithelial cells as part of host defense against fungal infection. Defensins, cathelicidins,
and histatins are important in preventing oral cavity [16]. Apart from antimicrobial activity, human
AMPs such as cathelicidins and defensins possess other functions such as immune modulation,
apoptosis, and wound healing [1–6]. It is now recognized that certain defensins also play a critical role
in sperm fertilization [17–19]. By the time this manuscript was completed, 103 human AMPs were
found in the APD [7,8]. A selected set of these peptides is provided in Table 1. The lengths of these
human peptides range from 10 (neurokinin A) to 149 amino acids (RegIIIα). Their net charges vary
from −3 (β-amyloid peptide) to +20 (antimicrobial chemokine CXCL9). On average, human AMPs
have 55 amino acids with a net charge of +5.6. Thus, both the average length and net charge for the
current list of human AMPs are higher than those averages from all the AMPs (32.4 residues and net
charge +3.2). An important reason for this is the inclusion of human antimicrobial proteins (> 100
amino acids) with high net charges (on average +10). The sequence diversity of human AMPs directly
determines their structural and functional diversity. This review article highlights the discovery, activity,
structure, mechanism of action, and therapeutic strategies of human host defense peptides selected
from the APD.
Table 1. Discovery timeline of select human antimicrobial peptides and proteins 1.

Year Name Sequence Source Activity 2 Ref.
1922 Lysozyme KVFERCELARTLKRLGMDG saliva, tears, G, F [20]
YRGISLANWMCLAKWESGY intestine
NTRATNYNAGDRSTDYGIFQ
INSRYWCNDGKTPGAVNAC
HLSCSALLQDNIADAVACAK
RVVRDPQGIRAWVAWRNRC
QNRDVRQYVQGCGV
1985 α-Defensin ACYCRIPACIAGERRYGTCIY Neutrophils, G, V, F, P, C [21]
HNP-1 QGRLWAFCC bone marrow
1985 α-Defensin CYCRIPACIAGERRYGTCIYQ Neutrophils, G, V, F, C [21]
HNP-2 GRLWAFCC bone marrow
1985 α-Defensin DCYCRIPACIAGERRYGTCIY Neutrophils, G, V, F, C [21]
HNP-3 QGRLWAFCC bone marrow
1988 Histatin 1 DSHEKRHHGYRRKFHEKHH saliva F [22]
SHREFPFYGDYGSNYLYDN
1988 Histatin 3 DSHAKRHHGYKRKFHEKHH saliva G, F [22]
SHRGYRSNYLYDN
1989 α-Defensin VCSCRLVFCRRTELRVGNCLI neutrophils G, V, F [23]
HNP-4 GGVSFTYCCTRV
1990 RNase 2 KPPQFTWAQWFETQHINMTS eosinophils V, P [24]
QQCTNAMQVINNYQRRCKN
QNTFLLTTFANVVNVCGNPN
MTCPSNKTRKNCHHSGSQVP
LIHCNLTTPSPQNISNCRYAQ
TPANMFYIVACDNRDQRRDP
PQYPVVPVHLDRII
1990 RNase 3 RPPQFTRAQWFAIQHISLNPP neutrophils G, V, P [24]
(Eosinophil RCTIAMRAINNYRWRCKNQ
cationic protein, NTFLRTTFANVVNVCGNQSI
ECP) RCPHNRTLNNCHRSRFRVPL
LHCDLINPGAQNISNCTYAD
RPGRRFYVVACDNRDPRDSP
RYPVVPVHLDTTI
1992 α-Defensin ATCYCRTGRCATRESLSGVC Paneth G, V, F [25]
HD-5 EISGRLYRLCCR cells/intestine,
female
reproductive
system
Table 1. Cont.
1993 α-Defensin AFTCHCRRSCYSTEYSYGTC Paneth V, F [26]
HD-6 TVMGINHRFCCL cells/intestine
1995 β-Defensin DHYNCVSSGGQCLYSACPIF Kidney, Skin, G, F, C [27]
hBD-1 TKIQGTCYRGKAKCCK salivary glands
1995 Cathelicidin LLGDFFRKSKEKIGKEFKRIV neutrophils; skin G, V, F, P, C [28–30]
LL-37 QRIKDFLRNLVPRTES
1997 β-Defensin GIGDPVTCLKSGAICHPVFCP skin, lung, G, V, F [31]
hBD-2 RRYKQIGTCGLPGTKCCKKP epithelia, uterus,
salivary glands
1998 Granulysin GRDYRTCLTIVQKLKKMVD cytolytic T and G, F, P, C [32]
KPTQRSVSNAATRVCRTGRS NK cells
RWRDVCRNFMRRYQSRVTQ
GLVAGETAQQICEDLR
1999 Ubiquicidin KVHGSLARAGKVRGQTP macrophages G [33]
KVAKQEKKKKKTGRAK
RRMQYNRRFVNVVPTFG
KKKGPNANS
2000 Thrombocidin-1 AELRCMCIKTTSGIHPKNIQS human blood G, F [34]
(TC-1) LEVIGKGTHCNQVEVIATLK platelets
DGRKICLDPDAPRIKKIVQKK
LAGDES
2000 Hepcidin 25 DTHFPICIFCCGCCHRSKCGM plasma, G, F [35]
(LEAP-1) CCKT Urine/Liver
2000 Neuropeptide SYSMEHFRWGKPV brain G+, V, F [36]
α-MSH
2001 β-Defensin GIINTLQKYYCRVRGGRCAV Skin, salivary G, V, F [37]
hBD-3 LSCLPKEEQIGKCSTRGRKCC glands
RRKK
2001 β-Defensin FELDRICGYGTARCRKKCRS testis, lung, G [38]
hBD-4 QEYRIGRCPNTYACCLRKWD kidney,
ESLLNRTKP neutrophils
2001 Dermcidin SSLLEKGLDGAKKAVGGLG eccrine G, F [39]
KLGKDAVEDLESVGKGAVH sweat/skin
DVKDVLDSV
2002 RNase 7 KPKGMTSSQWFKIQHMQPS urinary tract; G, F [40]
PQACNSAMKNINKHTKRCK respiratory tract;
DLNTFLHEPFSSVAATCQTP skin
KIACKNGDKNCHQSHGAVS
LTMCKLTSGKYPNCRYKEK
RQNKSYVVACKPPQKKDSQ
QFHLVPVHLDRVL
2003 RNase 5 QDNSRYTHFLTQHYDAKPQ Liver, skin, G+, F [41]
(angiogenin) GRDDRYCESIMRRRGPTSPC intestine
KDINTFIHGNKRSIKAICENK
NGNPHRENLRISKSSFQVTTC
KLHGGSPWPPCQYRATAGF
RNVVVACENGLPVHLDQSIF
RRPRP
Table 1. Cont.
2003 Chemokine SNFDCCLGYTDRILHPKFIVGF skin G, F, P [42]
CCL20 TRQLANEGCDINAIIFHTKKKL
SVCANPKQTWVKYIVRLLSK
KVKNM
2003 Chemokine TPVVRKGRCSCISTNQGTIHL blood G, P [42]
CXCL9 QSLKDLKQFAPSPSCEKIEIIAT
LKNGVQTCLNPDSADVKELIK
KWEKQVSQKKKQKNGKKHQ
KKKVLKVRKSQRSRQKKTT
2005 Psoriasin MSNTQAERSIIGMIDMFHKYT Skin, salivary G- [43]
(S100A7) RRDDKIDKPSLLTMMKENFPN glands, breast
FLSACDKKGTNYLADVFEKK
DKNEDKKIDFSEFLSLLGDIAT
DYHKQSHGAAPCSGGSQ
2006 RegIIIα EEPQRELPSARIRCPKGSKAY intestine G+ [44]
GSHCYALFLSPKSWTDADLA
CQKRPSGNLVSVLSGAEGSFV
SSLVKSIGNSYSYVWIGLHDP
TQGTEPNGEGWEWSSSDVMN
YFAWERNPSTISSPGHCASLSR
STAFLRWKDYNCNVRLPYVC
KFTD
2008 Substance P RPKPQQFFGLM the nervous G, F [45]
system
2008 Drosomycin- CLAGRLDKQCTCRRSQPSRRS oral epithelial F [46]
like defensin GHEVGRPSPHCGPSRQCGCH cells, skin
(DLD) MD
2009 Elafin AQEPVKGPVSTKPGSCPIILIR γδ T cells G, F, V [47]
CAMLNPPNRCLKDTDCPGIKK
CCEGSCGMACFVPQ
2010 β-amyloid DAEFRHDSGYEVHHQKLVFF brain G, F [48]
peptide 1-42 AEDVGSNKGAIIGLMVGGVVI
2011 Chemerin ELTEAQRRGLQVALEEFHKHP skin G, F [49]
PVQWAFQETSVESAVDTPFPA
GIFVRLEFKLQQTSCRKRDWK
KPECKVRPNGRKRKCLACIKL
GSEDKVLGRLVHCPIETQVLR
EAEEHQETQCLRVQRAGEDP
HSFYFPGQFAFS
2012 Amylin KCNTATCATQRLANFLVHSS pancreatic G [50]
NNFGAILSSTNVGSNTY β-cells
2012 KDAMP RAIGGGLSSVGGGSSTIKY eyes G- [51]
2013 DEFB114 DRCTKRYGRCKRDCLESEKQI epididymis G, F [19]
DICSLPRKICCTEKLYEEDDMF
1
Data from the APD [7,8]. For a complete list of human AMPs, please visit the APD website
(http://aps.unmc.edu/AP) and search in the name field using ―human‖. 2 In the APD, antimicrobial activities
against different types of microbes are annotated as below: G, bacteria; G+, Gram-positive bacteria only;
G-, Gram-negative bacteria only; F, fungi; V, viruses; P, parasites; C, cancer cells.
2. Identification of Human Antimicrobial Peptides
Antimicrobial substances might have been noticed long time ago [1–6]. Human lysozyme
(130 amino acids), discovered in saliva by Alexander Fleming in 1922 [20], is recognized as the first
antimicrobial protein [6]. However, the isolation and characterization of many more AMPs with
defined amino acid sequences did not start until the 1980s (please refer to the annual AMP discovery
plot in ref. [52]). Two major methods were utilized for AMP identification. Initially, chromatographic
approaches were used to isolate and characterize new peptides. With the recognition of peptide
sequence motifs, bioinformatic approaches were later developed to identify AMPs at the genomic
level. In the following, we describe the discovery of the major families of human antibacterial peptides
identified during 1985 and 2013.
2.1. Human Defensins
Host defense peptides are usually expressed as precursor proteins and the mature form is released
by protease processing. In 1985, the Lehrer group isolated a family of the mature form of α-defensins
from human blood [21]. Based on the source, property and size, these peptides were named as human
neutrophil peptides (HNP-1, HNP-2, and HNP-3). The three defensins have nearly identical amino
acid sequences (Table 1). Compared to HNP-2, both HNP-1 and HNP-3 contain only one additional
amino acid residue at the N-terminus: alanine for HNP-1 and aspartate for HNP-3. This additional
acidic residue in HNP-3 may make HNP-3 less active than HNP-1 or HNP-2 in killing Staphylococcus
aureus, Pseudomonas aeruginosa, and Escherichia coli. A fourth human neutrophil defensin, HNP-4,
was reported in 1989 [23]. It was also purified to homogeneity by chromatographic methods. HNP-4
has a distinct peptide sequence with 33 amino acids. In vitro, purified HNP-4 was shown to kill E. coli,
Streptococcus faecalis, and Candida albicans. Lehrer and colleagues found that HNP-1, HNP-2, and
HNP-3 are abundant in bone marrow, and can be detected in peripheral blood leukocytes, spleen and
thymus by RT-PCR [53].
In 1989, Ouellette and colleagues identified α-defensin genes from mouse Paneth cells [54]. Bevins
hypothsized that such orthologs also exist in human epithelial cells as part of the host defense
mechanism. A genetic approach was developed to map additional defensin genes based on the high
conservation of the nucleotide sequences in the signal coding region as well as the untranslated 5′
region. Using the probes from the conserved regions, they cloned the gene of HD-5 from human Paneth
cells [25]. Likewise, HD-6 was identified [26]. These peptides are tissue-specific as they are only
expressed in the Paneth cells of human intestines. The six human α-defensins (Table 1) share the same
disulfide bond pattern. If we number the six cysteines in Roman numbers: I, II, III, to VI, the three
disulfide bonds in α-defensins are CI–CVI, CII–CIV, and CIII–CV. Interestingly, alpha defensins have
been found in the neutrophils of rabbits, rats, hamsters, and guinea pigs, but not in mice or pigs [55].
Members from the human β-defensin family were discovered in the 1990s. Different from
α-defensins, the three disulfide bonds in β-defensins are CI–CV, CII–CIV, and CIII–CVI. Also,
β-defensins have a slightly longer sequence to allow for an additional helical region. The first human β
defensin (hBD-1) was reported by Bensch et al. from human plasma in 1995 [27]. Quantitative mRNA
analysis revealed kidney as the major source for hBD-1 [56]. In addition, different truncated forms of
hBD-1 were also isolated and found to be active against E. coli. The activity of hBD-1 might have been
compromised in cystic fibrosis (CF) lung due to its salt-sensitive activity against P. aeruginosa [57].
Subsequently, hBD-2 was identified from lesional psoriatic skin using the whole E. coli affinity
column [32]. This material was selected for AMP isolation based on the fact that patients with lesional
psoriatic skin have fewer skin infections than expected. This peptide is effective in killing
Gram-negative bacteria E. coli, P. aeruginosa, and yeast C. albicans, but is only bacteriostatic against
Gram-positive S. aureus. Like hBD-1, the activity of HBD-2 is also salt-sensitive [58]. In 2001, both
hBD-3 and hBD-4 were documented [37,38]. Based on this disulfide bond linkage pattern, hBD-3 was
also identified by using a bioinformatic approach [59,60]. In contrast to hBD-1 and hBD-2, hBD-3
remained active against S. aureus and vancomycin-resistant Enterococcus faecium at physiological salt
concentrations [37]. Bioinformatic studies led to the identification of 28 additional human and 43
mouse β-defensin genes in the respective genome [61]. Several of these β-defensins (hBD-6, hBD-26,
hBD-27, hBD-28, and DEFB114) are indeed antimicrobial in vitro [62–64]. This sequence motif-based
peptide prediction may be applied to any other species with a completed genome.
In insects, the activation of TOLL directly leads to the expression of drosomycin against fungal
infection [65]. In 2008, human drosomycin-like defensin was detected in oral mucosa [46,66],
indicating that this ancient innate defense mechanism is conserved. Sequence alignment in the APD
revealed 40% similarity to insect drosomycin, which comprises one α-helical and three β-strands.
Therefore, such a combined structure resembles human β-defensins to some extent. This peptide
appears to be specifically effective against filamentous fungi (e.g., Aspergillus spp) as it did not kill
tested yeast, Gram-positive or Gram-negative bacteria. Although the connection pattern of the six
cysteines has not yet elucidated, the resulting three disulfide bonds are critical for the antifungal
activity of human drosomycin-like defensing [46].
It is interesting to mention that a different type of defensins (called θ-defensins) has been identified
from non-human primates [66,67]. These 18-residue defensins are circular due to the formation of a
peptide bond between the N- and C-terminal ends. Like α- and β-defensins, they are also stabilized by
three sets of disulfide bonds (CI–CVI, CII–CV, and CIII–CIV). These defensins are generated by
liganding two truncated α-defensins. These genes are not expressed in humans due to the existence of a
premature stop codon. Like monkey θ-defensins, synthetic peptides corresponding to these human
counterpart pseudogenes are HIV-1 inhibitory [68–70]. It is uncertain whether the loss of θ-defensins
made humans generally more susceptible to HIV-1 infection.
2.2. Human Histatins: Two Genes Multiple Peptides
Histatins are a family of AMPs rich in histidines. In 1988, histatins 1, 3, and 5 were isolated from
human saliva by size-exclusion chromatography followed by HPLC separation [22]. All three histatins
exhibit the ability to kill the pathogenic yeast, C. albicans. Subsequently, other histatins were also
isolated [71]. However, only histatins 1 and 3 are gene encoded [22,72], since others are the cleaved
products of these two peptides. The histatin genes are located on chromosome 4, band q13 and
exclusively expressed in human salivary secretions [73].
2.3. Human Cathelicidins: One Gene Multiple Peptides
The significance of cathelicidins in protecting humans from infection is established by data from
animal models [74–76]. Cathelicidin peptides were first isolated in 1989 [77]. The precursor proteins
of cathelicidins share a highly conserved N-terminal ―cathelin‖ domain, but have drastically different
antimicrobial sequences, ranging from Pro- and Arg-rich peptides, helical peptides, to disulfide-linked
sequences [78]. The word ―cathelicidin‖ was originally used to refer to the entire precursor protein.
However, it is now accepted as the family name for mature AMPs from the C-terminal region. Another
term hCAP-18 is abbreviated from human cationic protein of 18 kDa, representing the precursors prior
to the release of cathelicidin peptides. However, these terms are interchangeably used in the literature.
Based on the conserved sequences in the cathelicidin precursors, Agerberth and colleagues cloned
the only human cathelicidin gene and predicted the antimicrobial peptide as FALL-39 in analogy to
PR-39 discovered in cattle [28]. Using the probes designed based on rabbit CAP-18, Larrick et al. also
cloned the C-terminal antimicrobial peptide [29]. In the same year, Cowland et al. isolated a 19 kDa
precursor protein hCAP-18 from human neutrophils [30]. Subsequently, the European group isolated
the natural form of the mature human cathelicidin peptide from neutrophils [79]. Since the isolated
peptide contains 37 amino acids and starts with a pair of leucines, it was named LL-37, which is two
residues shorter than the initially predicted peptide FALL-39 [28]. Interestingly, ALL-38, another form
of human cathelicidin peptides, was also characterized in 2003 [80]. This alterative form contains one
more alanine at the N-terminus than LL-37. ALL-38 was generated in female vagina due to the action
of gastricsin on sperm hCAP-18. Antimicrobial assays revealed a similar activity spectrum for LL-37
and ALL-38 against a panel of bacteria, including E. coli, S. aureus, P. aeruginosa, and Bacillus
megaterium. Thus, ALL-38, as well as other active peptides such as SgI-29 [81], plays a defense role
in the human reproductive system. In addition, LL-37 fragments were isolated from human skin
by chromatographic approaches [82]. These fragments have varying activities compared to intact
LL-37. Hence, a single human cathelicidin gene has been processed into different forms of active
peptides [83]. This phenomenon, however, is not unique to humans. There is precedence that multiple
AMPs are programmed in a single plant gene [84]. One possibility for this is that different cathelicidin
fragments provide a means to expanding the functional space of the single human cathelicidin gene.
A different model is utilized by sheep, horses, and cattle, which produce multiple cathelicidins with
varying functions [85–88].
2.4. Human Dermcidin
Besides human cathelicidin LL-37 [89], dermcidin, an anionic defense peptide, was found in human
sweat [39]. Unlike human defensins and cathelicidins that are induced under inflammatory and injured
conditions, dermcidin is constitutively expressed in human sweat [90]. Furthermore, dermcidin
variants as well as fragments were also detected. It appears that the level of dermcidin did not vary
between healthy people and infected patients [91]. In addition, dermcidin may be related to other
human diseases such as cancer and atherosclerosis [92,93].
2.5. Human Hepcidins
Human liver expressed antimicrobial peptide-1 (LEAP-1) was discovered from human blood
ultrafiltrate in 2000 [35]. The same peptide was also found by Ganz et al. from human urine and
named as hepcidin 25 [94]. This liver-synthesized peptide is especially rich in cysteines (32%), leading
to four disulfide bonds in a 25-residue peptide. In 2009, the connection pattern of the four disulfide
bonds was revised to C7–C22, C10–C13, C11–C19, and C14–C22 [95]. This antimicrobial peptide
also plays an important role in ferrous use [96] and single-residue mutations in this molecule are
associated with severe juvenile hemochromatosis, a genetic disease of severe iron overload [97]. Unlike
LEAP-1, antimicrobial peptide LEAP-2, however, is not involved in the regulation of iron use [98].
2.6. Human AMPs Derived from Known Proteins
Some AMPs are derived from known proteins. Park et al. isolated buforin I from amphibians in
1996 [99]. Sequence comparison revealed that buforin I is a cleaved fragment of histone H2A. Of
interest, this mRNA was also detected in humans, suggesting a possible role of this peptide in
antimicrobial defense [100].
The human airways are essential for the exchange of molecules with the environment. It is
necessary to guard this channel to prevent the infection of microbes in the air. In 2001, Ganz and
colleagues isolated calcitermin primarily targeting Gram-negative bacteria. This 15-residue peptide is
derived from the C-terminus of calgranulin C (a S100 protein). It contains three histidines (His9,
His11, and His13) at the N-terminus and has the potential to adopt a helical conformation in
membranes. These histidines may explain its enhanced activity in acidic buffers (pH 5.4) and in the
presence of micromolar concentrations of ZnCl2 [101].
Another example for protein-derived AMPs is KDAMP, keratin-derived AMPs, which were
identified from bactericidal lysate fractions of human corneal epithelial cells. These molecules are rich
in glycines [51]. The glycines appear to be important for killing P. aeruginosa as substitution of a
string of glycines with alanines reduced peptide potency. A search of the APD reveals that glycine-rich
(>25%) AMPs have also been identified in bacteria, plants, insects, spiders, nematodes, crustaceans,
fish, and amphibians. Thus, glycine-rich peptides constitute a common molecular design for host
defense [102–110].
2.7. Antimicrobial Chemokines and AMPs from Human Immune Cells
The fact that some AMPs possess chemotactic effects inspired the evaluation of antimicrobial
activity of chemokines, which are known for chemotaxis. In 2000, Krijgsveld et al. found antimicrobial
thrombocidins, peptides derived from CXC chemokines in human blood platelets [34]. In 2003,
Oppenheim and colleagues identified 20 antimicrobial chemokines [42] and additional two members
(CCL27 and CCL28) were also reported by Hieshima and colleagues [111]. In 2011, antimicrobial
activity of chemotactic chemerin was also reported [49]. Further studies are required to establish the
in vivo relevance of the in vitro activity of these chemokines. The common nature of chemokines and
AMPs, however, bridges the innate and adaptive immune systems.
Antimicrobial peptides have also been found in other immune cells. In 1999, Hiemstra et al.
identified ubiquicidin from ribosomal protein S30 in various tissues. This peptide is active against
Listeria monocytogenes, S. aureus, Salmonella typhimurium, and E. coli. Considering the fact that
L. monocytogenes can live within cells, the expression of ubiquicidin in macrophages would limit the
replication of this bacterium. Interestingly, ribosomal protein S30 is identical in rats, mice and
humans [33]. In 1998, granulysin (74 amino acids) was detected in human cytotoxic T cells and natural
killer (NK) cells [32]. Similar proteins called NK-lysins are found in other animals [112]. Granulysin
is active against Gram-positive and Gram-negative bacteria, and fungi, including mycobacteria.
Thus, the human adaptive immune system has incorporated an innate defense molecule for direct
disruption of tumor cells or invading microbes. In addition, human γδ T cells produce antimicrobial
peptide elafin [47]. Like human secretory leucoprotease inhibitor (SLPI), elafin was initially identified
as a protease inhibitor. Elafin shows an inhibitory effect on bacteria, fungi, and viruses [113].
2.8. Antimicrobial Neuropeptides
Antibacterial peptides were also isolated from the neuroendocrine system from cattle. Secretolytin
corresponds to the C-terminal fragment (residues 614–626) of bovine chromogranin B [114]. This
peptide displayed activity against M. luteus and reduced the growth of B. megaterium. Vasostatin-1 is
a 76-residue N-terminal fragment of bovine chromogranin A. This peptide is active against both
Gram-positive bacteria and fungi [115]. It is conserved in humans, pigs, horses, mice, rats, and frogs.
Catestatin is a 21-residue AMP derived from human chromogranin A [116]. Subsequently, cattle
enkelytin, the C-terminal fragment corresponding to residues 209–237 of proenkephalin-A was also
found to be antibacterial [117]. A similar proenkephalin system exists in humans although the
processing machinery can differ [118]. Thus, these neuropeptides also play a communication role
between neuroendocrine and the immune system [119].
In 1998, human neuropeptide Y (36 residues) was demonstrated to have antimicrobial activity [120].
Interestingly, the antifungal activity of this peptide against C. albicans increased several folds by
truncating N-terminal 12 residues, implying the importance of the helical region 14–32. A more recent
study, however, only observed activity against E. coli, but not C. albicans [121]. Future studies will
clarify whether the peptide has direct antimicrobial effects. In 1999, adrenomedullin, usually expressed
on surface epithelial cells, was reported to have antibacterial activity against all the tested bacteria,
but not C. albicans [122]. Alpha-MSH was added to the human AMP list in 2000 as well [36]. These
findings extended host defense peptides to the nervous system [123]. Since then, more antimicrobial
neuropeptides have been documented [45,124]. Some of these neuropeptides may become leads for
developing new antibiotics. For example, alarin is a human brain neuropeptide with activity against
only Gram-negative bacteria such as E. coli, but not Gram-positive bacteria such as S. aureus [124].
In particular, α-MSH showed in vitro antifungal activity against C. albicans at fM to pM [125],
much lower than nM for some bacterial lantibiotics and µM for many cationic peptides.
2.9. Beta-Amyloid Peptides
Beta-amyloid peptides have long been thought to be the culprit of Alzheimer's disease. It is believed
that the β-sheet form, not the helical form of the peptide, is toxic. A recent demonstration of
antimicrobial activity for β-amyloid peptides adds a new research dimension to this worrisome
disease [48]. Further studies along this line could be useful to provide novel insight into the
mechanism of this human disease. In 2012, amylin (human islet amyloid polypeptide, hIAPP) was
found bactericidal, too [50]. Transformation of amylin into toxic fibrils can disrupt cell membranes
and lead to β-cell death, perhaps one of the causative factors of type 2 diabetes mellitus. The aggregated
form of amylin is likely to exert its cytotoxicity by damaging cell membranes [126]. It is noticeable
that, when in excess, bacteria microcin E492 can form fibrils as a storage form and lose antimicrobial
activity [127]. Is there anything in common here from bacteria to humans? Perhaps, this is one
mechanism that nature attempts to remove the toxic effects of an over expressed protein, including
antimicrobial peptide.
2.10. Human Antimicrobial Proteins
There are 14 antimicrobial proteins (>100 amino acids) in the APD database as of March 2014 [8].
This includes the first antimicrobial protein lysozyme [20]. Several eosinophil proteins were purified
in 1990. Both eosinophil-derived neurotoxin (EDN, or RNase 2) and eosinophil cationic protein (ECP,
or RNase 3) possess anti-parasitic activity against Brugia pahangi and Brugia malayi [24]. These two
ribonucleases are also active against respiratory syncytial virus (RSV) with a single-stranded
RNA [128]. In addition, RNase 3 has a unique bacterial agglutinating activity [129]. In 2002, RNase 7
was identified from human skin [40]. Remarkably, it is active against bacteria such as Mycobacterium
vaccae and yeast, even at 4 °C [130]. Recombinant RNase 7 exhibited antimicrobial activity against
uropathogens (E. coli, P. aeruginosa, Klebsiella pneumonia, Proteus mirabilis, E. faecalis, and
Staphylococcus saprophyticus) [131,132]. In fact, RNase 7 is the most abundant innate defense
peptide in the human urinary tract [132], although the contributions of other human AMPs cannot be
ignored [56,133–135]. Human RNase 5, (also known as angiogenin or ANG) with weak RNase
activity, is initially implicated in angiogenesis. It appears that the nucleus location is necessary for
angiogenesis [136]. In 2003, RNase 5 was found to have a toxic effect on Gram-positive bacteria and
fungi [41]. However, another study found little activity using a commercial material [137]. In 2006,
RNase 8 was found to inhibit M. vaccae [138,139]. Therefore, of the eight human ribonucleases, RNases
2, 3, 5, 7, and 8 appear to play a role in host defense.
Psoriasin is another human protein with multiple functions. It is identical to S100A7, a protein
member of the S100 family. Psoriasin was initially characterized as a Ca2+ binding protein with
chemotactic property from human skin psoriatic lesions [140,141]. Subsequently, psoriasin was found
in cancer lesions, making it a potential cancer biomarker [142]. However, the antimicrobial activity of
psoriasin against E. coli was not demonstrated until 2005 [43].
Three RegIII (regenerating gene family protein III) proteins were initially identified in mice [143].
RegIIIα, a RegIIIγ homolog protein, was found in humans. Different from RegIIIβ from mice, human
RegIIIα only inhibited the growth of Gram-positive bacteria such as L. monocytogenes, Listeria
innocua, and E. faecalis [44].
3. Antimicrobial and Anticancer Activities of Human Antimicrobial Peptides
3.1. Antibacterial Activities
Antibacterial activities of human AMPs have been mentioned in the preceding section. This section
provides an overview of the activity spectrum of these peptides. Based on the APD [8], the majority of
human AMPs (90 out of 103) can inhibit the growth of bacteria. They display a broad-spectrum
activity against a variety of Gram-positive and Gram-negative bacteria. However, three human AMPs
kill primarily Gram-positive bacteria. RNase 5 has an effect on S. pneumonia [41], while α-MSH is
inhibitory to S. aureus [36]. A third AMP, RegIIIα, is active against L. monocytogenes, L. innocua,
and E. faecalis [44]. In addition, ten human AMPs are inhibitory mainly to Gram-negative bacteria.
These include hBD-26, hBD-27, human calcitermin, psoriasin/S100A7, CCL8, CCL13, CCL19, alarin,
HMGN2, and KDAMP peptides [42,43,51,63,101]. They may control different Gram-negative pathogens.
For example, both calcitermin and HMGN2 were active against E. coli and P. aeruginosa [101,144].
Alarin is active against E. coli, while the KDAMP peptides are primarily active against P. aeruginosa.
Thus, these peptides may form the basis for developing new antimicrobials with a desired antibacterial
activity spectrum.
3.2. Antiviral Activity
Of the 103 human AMPs, 16 are virucidal. They include the six well-characterized human
α-defensins (HNP-1, HNP-2, HMP-3, HNP-4, HD-5, and HD-6), three β-defensins (hBD-1, hBD-2,
and hBD-3), cathelicidin LL-37, histatin 5, α-MSH, elafin, SLPI, CXCL12, RNase 2, and RNase 3.
Elafin is the major antiviral protein in cervicovaginal lavage fluid [113]. Both RNase 2 and RNase 3
inhibit respiratory syncytial viruses (RSV) [145,146]. SLPI levels in saliva and semen, but not breast
milk, approximate the level required for HIV-1 inhibition in vitro [147]. Although HNP-1 is a lectin
that binds to gp120 and CD4, it shows an inhibitory effect after HIV-1 entry [148]. It also inhibits
non-enveloped BK virus infection by aggregating virions and blocks binding to host cells [149]. Of the
four human β-defensins, hBD-2 and hBD-3 can be induced by viral infection and block HIV replication
by directly neutralizing the virions and through modulation of the CXCR4 coreceptor [150]. Human
cathelicidin LL-37 is also inhibitory to HIV-1 [151,152]. Wang et al. further dissected the active
region of LL-37. While the central fragment GI-20 showed an optimal therapeutic index, the LL-37
core peptide FK-13 contains the minimal anti-HIV sequence [152]. In the histatin family, only a
histatin-5-derived peptide has an effect on HIV-1 [153]. For a systematic review of AMPs with known
anti-HIV activity, interested readers may refer to a recent review article [154].
3.3. Antifungal Activity
In the APD, 58 human AMPs are fungicidal [8]. Typical examples are human α-defensins,
cathelicidin LL-37, hepcidins, and histatins. In the case of LL-37, protease processing into fragments
such as KS-30 and RK-31 is essential in inhibiting C. albicans [155]. In addition, the antifungal
activity of LL-37 depends on both media and pH. Both LL-25 and RK-31 can rapidily enter the cell
cytoplasm [156]. It is likely that these LL-37 fragments target intracellular molecules. It appears that
such cathelicidin fragments in human sweat play a role in human skin innate defense against fungal
infection [155].
3.4. Antiparasitic Activity
Some human AMPs also have antiparasitic properties. These include HNP-1, LL-37, granulysin,
CCL2, CCl20, CCL28, CXCL4 (hPF4), CXCL6, CXCL9, CXCL10, RNase 2, and RNase 3. RNases 2
and 3 might be the earliest AMP examples from humans that were demonstrated to have antiparasitic
activity [24]. Other more recent examples are HNP-1 against the promastigotes and amastigotes forms
of Leishmania major [157], chemokine CCL28 against Leishmania mexicana [158], LL-37 against
Entamoeba histolytica [159], and Platelet factor 4 (hPF4) against malarial parasite Plasmodium
falciparum [160]. These examples verify that human defense peptides also play a role in controlling
parasite infections.
3.5. Anticancer Activity
There is a growing interest in developing AMPs into anticancer peptides. Magainins, cecropins, and
defensins were all shown to have anticancer effects [161–163]. Many other AMPs possess this activity
as well and some were discussed in previous review articles [164–166]. A more complete and updated
list of anticancer AMPs can be searched in the APD database [8]. The 166 anticancer peptides cover
multiple sources, including animals (105 peptides), plants (48 AMPs), bacteria (seven bacteriocins),
fungi (one peptide), and laboratory synthesis (five peptides).
The anticancer activities of human AMPs have not been widely evaluated since only six members
are annotated as anticancer in the APD [8]. They are HNP-1, HNP-2, HNP-3, hBD-1, LL-37, and
granulysin. Indeed, reduced expression of granulysin in patients is correlated with the progression of
cancer [167]. However, the level of granulysin increased substantially in cancer cells [168]. Similar
observations have been made with human defensins and LL-37. While HNP-1 inhibits the growth of
human lung adenocarcinoma xenograft in nude mice [169], the same molecule can be overexpressed in
tumors [170,171]. HBD-1 can suppress urological and prostate cancers [172,173]. In oral squamous
cell carcinoma, hBD-1 also suppressed tumor proliferation, whereas hBD-2 and hBD-3 showed an
opposite effect [174]. Likewise, human cathelicidin LL-37 is overexpressed in breast, ovarian, and
lung cancers, but it suppresses tumorigenesis in gastric cancer [175]. Such a complex involvement of
AMPs in various cancers deserves additional studies. In particular, it is important to elucidate the
factors that trigger the overexpression of AMPs in certain cancer cells.
Our current knowledge, however, may be utilized to our advantage. For instance, the over-expression
of human cathelicidin LL-37 or defensins may serve as useful biomarkers for cancer diagnosis [176–178].
In addition, the fragments of LL-37 with demonstrated anticancer effects in vitro [179] and in vivo [180]
might constitute useful templates for designing new anti-tumor drugs, especially those resistant to
existing therapeutics. Anticancer AMPs can work by different mechanisms. The effects may result
from a direct bacterial killing when bacteria could be the culprit of cancer (e.g., gastric cancer) [175].
AMPs may selectively kill cancer cells in part due to exposed anionic phosphatidylserines (PS) [181].
It is also possible that some AMPs kill cancer cells indirectly by inducing apoptosis (see below).
3.6. Cytotoxic Effects of Human AMPs
It is accepted that many AMPs target bacterial membranes. While bacteria are abundant in anionic
lipids such as phosphatidylglycerols (PG) and cardiolipin (CL) [83], human cells comprise zwitterionic
phosphocholines (PC) and cholesterol. The differences in membranes of bacterial and human cells to a
large extent determine cell selectivity of cationic AMPs. Indeed, among the 103 human AMPs, only
three peptides are annotated to have cytotoxic effects on mammalian cells. In the case of LL-37, we
found it possible to reduce the peptide cytotoxicity by decreasing peptide hydrophobicity [179,182].
The relatively low cytotoxicity of human AMPs makes them attractive templates for engineering
new antimicrobials.
In addition to membrane differences, human cells could use other mechanisms to reduce or remove
the potential toxic effect of AMPs on themselves. By expressing a peptide called p33 on the cell
surface, the toxic effects of human LL-37 is masked [183]. Instread of directly secreting AMPs,
humans also release exosomes (nanovesicles enriched in host defense peptides) into the urinary tract to
keep it sterile [184]. It may be speculated that such exosomes, similar to artificial AMP-containing
liposomes, could be an effective way to reduce cytotoxicity to human cells.
3.7. Other Biological Functions of Human AMPs
In addition to antimicrobial and anticancer activities, many human AMPs possess other functions
such as chemotaxis, apoptosis, and wound healing:
Chemotactic activity. Currently, 35 human AMPs have chemotactic properties. Examples are
defensins and cathelicidin LL-37. Antimicrobial chemokines were originally identified for chemotactic
activity. A clear difference between chemokines and AMPs is the concentration needed for action.
While the chemotactic effect needs peptides in the nM-pM range, antimicrobial action usually requires
µM peptides [185]. Another difference between chemotactic and antimicrobial activities lies in the
molecular target. While AMPs usually target membranes of invading bacteria, the chemotactic effects
require the association of peptides to host cell receptors. As one example, human LL-37 achieves its
chemotactic ability to monocytes, macrophages, neutrophils, and T cells by binding formyl peptide
receptor-like 1 (FPRL-1) [186].
Apoptosis. Apoptosis is the process of programmed cell death. Different factors can trigger cell
apoptosis. AMPs are one of these factors. Interestingly, apoptosis may be induced or suppressed by the
same peptide depending on the biological context. Human LL-37 induces apoptosis in vascular smooth
muscle cells, primary airway epithelial cells, oral squamous cell carcinoma SAS-H1 cells, intact rat
aorta rings and cultured rat aorta smooth muscle cells, Jurkat T-cells and A549 cells [187–191], while
it suppresses this process in keratinocytes and neutrophils [192,193]. As a consequence, elucidation of
the mechanism to control apoptosis may offer new therapeutic strategies. A recent interesting finding
is that inhibition of human LL-37-induced apoptosis by administrating urothelial glycosaminoglycan
(GAG) analogs can prevent the development of interstitial cystitis (IC) in a mouse model [194]. LL-37
could suppress the lipopolysaccharides (LPS)-induced apoptosis of endothelial cells, thereby
attenuating lethal sepsis/endotoxin shock [195]. In the case of colon cancer, however, activation of
apoptosis suppresses tumorigenesis [196]. It seems that apoptosis requires the major antimicrobial
region FK-16 (i.e., corresponding to residues 17-32) of human cathelicidin LL-37 [179,180].
Wound healing. Human host defense peptides can also promote wound healing, a process of injury
repairs. Salivary histatin 2 can enhance fibroblast cell migration, whereas human LL-37 at 1 µM can
induce cell migration and promote proliferation [197]. It is proposed that these peptides play a role in
fast wound coverage.
In summary, antimicrobial activity is a common property of human AMPs, although the in vivo
relevance has not firmly established for each polypeptide. Furthermore, human AMPs can perform
other functions depending on the biological context, peptide concentration, proteases, and the
metabolic state. Under diseased conditions, AMPs may have an opposite effect (e.g., cancer
suppression vs. progression). Understanding the elegant balance of AMPs in these processes in the
healthy state as well as the factors that could tilt the balance to a diseased state may yield useful means
for cancer treatment.
4. Three-Dimensional Structures of Human Antimicrobial Peptides
Three-dimensional structure of human host defense AMPs are helpful to understand the function of
AMPs described above. Many short and linear antimicrobial peptides do not have a folded structure
free in solution. However, they may become structured upon interactions with host cells by binding to
a specific receptor to trigger the biological responses to microbial invasion. In addition, such AMPs
may also adopt a defined structure upon association with bacterial targets such as membranes.
The bound structure in either category is not trivial to determine. Membrane-mimetic models are
normally utilized to determine the membrane-bound structures of receptors or AMPs. These models
include organic solvents, detergent/lipid micelles, lipid bicelles, nanodiscs, and lipid bilayers [198,199].
The known 3D structures of these short peptides are primarily determined by multi-dimensional
solution nuclear magnetic resonance (NMR) spectroscopy [199]. Some AMPs possess a folded
structure in aqueous solutions primarily due to the structural stabilization of disulfide bonds.
The structures of these small proteins can be determined by NMR or X-ray crystallographic methods.
When both methods are applied, similar structures are usually found. In addition, NMR measurements
can gain insight into protein dynamics (i.e., motions). Among the 103 human AMPs annotated in the
APD database, 42 have a known 3D structure (27 determined by NMR and 15 by X-ray) [8].
Although there are different classification schemes [1–6,200], the structures of natural AMPs fall
into four large families (α, β, αβ, and non-αβ) [201]. Peptides in the α family contain α-helical
structure as the major secondary structure. Typical examples are human cathelicidin LL-37, histatins,
dermcidin, and granulysin. The β family is characterized by at least a pair of two β-strands in the
structure. Human α-defensins, hepcidins, and SLPI use this type of structure. The αβ family contains
both α and β secondary structures, whereas the non-αβ family has neither α nor β structure (also called
extended structures). While there are multiple structural examples for the αβ family (Table 2),
no structural example has been found for the non-αβ family of human AMPs. In the following,
we highlight atomic structures of human AMPs from the α, β, and αβ families. These structures are
annotated in the APD database [8] and structural coordinates can be obtained from the Protein Data
Bank (PDB) [202] via the APD links.
Table 2. Properties of selected human antimicrobial peptides with known 3D structure 1.

APD ID Peptide name Length Net charge Pho% Boman index Structure class
2257 Lysozyme 130 +8 40 2.28 α
505 Histatin 5 24 +5 8 4.81 α
780 Lactoferricin 49 +10 36 3.14 α
310 LL-37 37 +6 35 2.99 α
433 Dermcidin 47 −2 38 1.11 α
1161 Granulysin 74 +11 33 3.5 α
2072 Psoriasin/S100A7 101 −1 32 2.3 α
β-Amyloid
1676 42 −3 45 0.77 α
peptide 1-42
176 HNP-1 30 +3 53 1.07 β
177 HNP-2 29 +3 51 1.17 β
178 HNP-3 30 +2 50 1.42 β
179 HNP-4 33 +4 51 1.4 β
180 HD-5 32 +4 40 2.6 β
181 HD-6 32 +2 40 1.71 β
192 Hepcidin 20 20 +3 60 0.46 β
Hepcidin 25
193 25 +2 52 0.89 β
(LEAP-1)
2095 SLPI 107 +12 34 1.87 β
451 hBD-1 36 +4 36 1.3 αβ
524 hBD-2 41 +7 36 0.9 αβ
283 hBD-3 45 +11 33 2.87 αβ
811 LEAP-2 40 +4 40 2.94 αβ
2067 RNase 5 125 +11 28 2.99 αβ
2073 RNase 7 128 +16 32 2.16 αβ
2071 RegIIIα 149 +1 33 1.77 αβ
2085 CCL1 73 +10 41 2.25 αβ
2086 CCL8 75 +6 37 2.27 αβ
2088 CCL13 75 +11 36 1.89 αβ
2075 CCL20 69 +8 43 1.34 αβ
2187 CCL27 56 +1 41 1.57 αβ
2076 CXCL1 73 +6 38 1.51 αβ
2080 CXCL10 77 +11 36 2.25 αβ
1
Obtained from the Antimicrobial Peptide Database (http://aps.unmc.edu/AP) [8]. Peptide hydrophobic
amino acid content (percent) is represented by pho% in the table. Protein-binding potential [1] was re-named
as Boman index in the APD database in 2003.
4.1. The α-Helical Family: Histatins, Cathelicidins, Dermcidin, and Granulysin
Histatins. Histatins 1, 3, and 5 are active against C. albicans and their candidacidal activities are in
the following order: histatin 5 > histatin 3 > histatin 1 [203]. Rai et al. investigated the relationship
between sequence length and activity using histatin 5 as the template. They found that the C-terminal
sequence is important [204]. P-113 with 12 residues corresponding to residues 4–15 of histatin 5
retained anti-candida activity [205]. Using histatin 3 as a model, Zuo et al. found increased activity
when the active sequence was expressed in tandem (repeated once) [206]. However, duplication of the
functional domain of histatin 5 did not enhance candidacidal activity [207]. These results suggest that
active domain duplication is not necessary a universal strategy for activity enhancement. Histatin 5 can
adopt a helical conformation in the presence of membrane-mimetic agents [204]. However, this helical
structure did not appear to be essential for candidacidal activity as peptides with a less helical structure
(achieved by proline insertion) can be equally active [208]. Histatin 5 contains a consensus sequence,
HEXXH, which is known to bind Zinc. Zinc binding can lead to vesicle fusion and helical
conformation as well [209]. Zinc binding actually potentiates peptide activity against gram-positive
bacteria E. faecalis [210]. An analog of histatin 5 was found to bind DNA and have nuclease activity
due to the synergistic oxidative and hydrolytic activities of the metal-peptide complex [211].
Human cathelicidin LL-37. To understand the structural basis of antimicrobial activity, 3D

triple-resonance NMR spectroscopy was used to solve a high-quality structure for human LL-37 in
the presence of sodium dodecyl sulfate (SDS) micelles [182]. LL-37 has a long helix covering
residues 2–31, whereas the C-terminal tail is disordered and does not superimpose well to each other
(Figure 1A). This structure is fully consistent with the backbone dynamics measured by an
independent NMR experiment. This amphipathic helical region determined in SDS micelles is
responsible for binding to bacterial outer and inner membranes. Interestingly, the LL-37 tail appeared
to be involved in peptide aggregation [212], which influences LL-37 activity [213]. The N-terminal
region of LL-37 is less important for antibacterial activity since LL-23, a natural fragment of LL-37, is
only active against susceptible E. coli or S. aureus strains. The weak activity of LL-23 is attributed to a
hydrophilic residue Ser9 that splits the hydrophobic face into two clusters [214]. This Ser9 residue also
segregates the hydrophobic surface of LL-37 into two hydrophobic domains. It is established that the
central helix (Figure 1B) of human LL-37 is critical for antibacterial, anti-biofilm, and antiviral
activity (reviewed in ref [83]).
Dermcidin. Unlike cationic cathelicidin LL-37 (net charge +6), dermcidin (DCD-1) has a net charge
of −2. This peptide also prefers anionic membranes, however. DCD-1L, a variant with one additional
leucine at the C-terminus, showed an enhanced affinity for membranes than DCD-1. In the membrane
bound state, DCD-1L has a helical conformation. It is located on the membrane surface and can
aggregate in the presence of zinc [215]. This oligomeric structure of dermcidin has recently been
determined by X-ray crystallography [216]. A hexameric helix-bundle structure (Figure 1C) is
proposed to insert into bacterial membrane as an ion channel. In the crystal, the Zn2+ coordinates
with a group of acidic amino acids. It should be pointed out that the 3D structure of DCD-1L was
also determined previously by 3D NMR spectroscopy using a 15N-labeled peptide in a 50%
trifluoenthanol (TFE) solution. Four helical regions were identified (α1: 5–7; α2: 10–12; α3: 26–33;
and α4: 36–45) [217]. In this case, TFE might have disrupted the oligomeric structure of dermcidin.
There are precedents for such an effect of TFE. For example, the oligomeric structure of a K+ channel
did not survive in TFE but retained in membrane-mimetic micelles such as SDS (reviewed
in ref. [198]). These examples emphasize the importance of determining the 3D structure of AMPs in a
proper environment.
Figure 1. Three-dimensional structures of human antimicrobial peptides from the α-helical

family: (A) and (B) human cathelicidin LL-37 determined by NMR spectroscopy (PDB ID:
2K6O); (C) dermcidin determined by X-ray crystallography (PDB ID, 2YMK); and (D)
granulysin determined by X-ray diffraction (PDB ID: 1L9L). In the case of LL-37, an
ensemble of five structures is shown to better view the disordered C-terminal tail (A),
whereas a space-filling model is given to show the segregation of the hydrophobic surface
(gold) into two domains (B) [182]. The longer one corresponds to the central helix which is
important for antimicrobial, anti-biofilm and antiviral activities [83]. Images were
generated by using the software MOLMOL [218]. Further details can be found in the text.
Granulysin. Incorporation of AMPs into cytotoxic T cells might have conferred the ability to lyse
cells. The crystal structure of granulysin is shown in Figure 1D. There are five helical regions (α1:
2–17; α2: 23–35); α3: 39–61; α4: 66–69; α5: 71–73) [219]. The sequences of human granulysin
(74 amino acids) and other NK-lysins share a low degree of similarity (~35%), but homologous
modeling reveals antimicrobial features (active helices and basic residue positions) are conserved [112].
Although granulysin only contains two disulfide bonds, it belongs to the saposin-like protein
family [220]. Saposin-like proteins comprise several helices usually stabilized by three disulfide
bonds [221]. Twelve AMPs in the APD [8] from amoebozoa, nematodes, and large animals such as
pigs are annotated to share the saposin-like protein fold [112]. To be antimicrobial, however, neither
the entire sequence nor the protein fold is needed. For example, synthetic peptides and analogs derived
from helices 3 and 4 are active against Vibrio cholera [222]. In addition, the peptide based on the
helix-bend-helix motif (residues 31–50) displayed similar antimicrobial activity against
Propionibacterium acnes (a key therapeutic target in acne) when synthesized entirely using D-amino
acids [223]. Because short peptides can be readily synthesized, they provide useful alternatives for
topical treatment of such bacterial infections.
4.2. The β Family: α-Defensins
Alpha-defensins. Structurally, α-defensins consist of three β-strands that form a β-sheet. In the
crystal, a dimeric structure is found for human HNP-1, where two copies of the molecule pack
together. HNP-2 and HNP-3 have a similar structure. Thus, it is primarily due to the single amino acid
difference in these defensins (Table 1) that influences peptide activity. With a more hydrophobic
sequence, HNP-4 is more potent against E. coli and C. albicans than other human α-defensins [23].
Using a kinetic 96-well turbidimetric procedure, the relative potencies of six human α-defensins were
compared. In the case of Gram-positive S. aureus, the activity is in the following order: HNP-2 >
HNP-1 > HNP-3 > HNP-4. In contrast, their relative potencies against Gram-negative E. coli is
HNP-4 > HNP-2 > HNP-1 = HNP-3 [224]. Thus, the antibacterial activities of these defensins are also
bacteria dependent. This likely reflects the distinct differences in membranes of these organisms. The
poor antibacterial activity of HNP-3 is not surprising considering the presence of an acidic aspartate at
the N-terminus of the peptide, making it unfavorable to target the negatively charged surface of
bacteria. HD-5 displayed a rather potent activity, which is comparable to HNP-2 against S. aureus and
HNP-4 against E. coli. The higher activities of HNP-4 and HD-5 against E. coli are correlated with
their higher net charge of +4 (Table 2). HD-6 has a poor antibacterial activity. In the crystal, it forms a
tetrameic structure (Figure 2B) [123].
Figure 2. Select 3D structures of human antimicrobial peptides from the β and αβ families:
(A) HNP-1 (dimeric crystal structure, PDB ID: 3GNY); (B) HD-6 (tetrameric crystal
structure, PDB ID: 1ZMQ); (C) hBD-3 (NMR structure, PDB ID: 1KJ6) and RegIIIα
(crystal structure, PDB ID: 4MTH). See the text for further details.
4.3. The αβ Family: β-Defensins, Antimicrobial Chemokines, RNases, and RegIIIα
Beta-defensins. Human β-defensins comprise both α and β structures in the same 3D fold.
Figure 2C shows the NMR structure of human β-defensin 3 (hBD-3), which starts with a helical
structure followed by three beta strands [225]. NMR translational diffusion studies revealed a dimer
for hBD-3, but a monomer for both hBD-1 and hBD-2 in solution. The stronger antibacterial activity
of hBD-3 than either hBD-1 or hBD-2 was attributed to the dimeric structure as well as higher charge
density on the protein surface [225]. Interestingly, the disulfide-linked form of hBD-1 is poorly active
and became highly potent against bacteria and fungus C. albicans under reduced conditions where the
disulfide-linked structure was disrupted [226]. It seems that the folded hBD-1 is the stored form, which
can be transformed into an active form when needed.
Antimicrobial chemokines. Chemokines interact with receptors to realize chemotactic functions.
They share a similar fold consisting of a three-stranded sheet followed by one α-helix at the
C-terminus [227–229]. The N-terminal region is frequently disordered (Figures 3A–C).
Figure 3. 3D structures of human chemokines with antimicrobial activity. Shown are (A)
CCL1 (NMR structure, PDB ID: 1EL0); (B) CCL8 (crystal structure, PDB ID: 1ESR); (C)
CCL11 (NMR structure, PDB ID: 2EOT); (D) CCL21 (NMR structure, PDB ID: 2L4N);
(E) CCL27 (NMR structure, PDB ID: 2KUM); (F) CXCL12 (NMR structure, PDB ID:
2KOL); (G) CCL20 (crystal structure, PDB ID: 1M8A); (H) CCL13 (crystal structure,
PDB ID: 2RA4); (I) CXCL1 (NMR structure, PDB ID: 1MSH); (J) CXCL10 (crystal
structure, PDB ID: 1O80).
This can be best seen using a superimposed structural ensemble for CCL20 or CCL27 [230,231]
determined by NMR (Figure 3D,E). The β-sheet appears to separate the N-terminal domain that
interacts with cell receptors and the C-terminal domain that contains the antimicrobial helix for
targeting bacterial membranes (Figure 3F) [232]. Some chemokines can also form oligomers.
The dimeric forms of CCL20, CCL13, CXCL1, and CXCL10 [233–236] are shown in panels G to J of
Figure 3. The dimer interface is normally composed of the C-terminal helix and strand 3. In the case of
CCL13, however, it is the N-terminal region that occupies the interface (Figure 3H). Yung et al. found
a direct binding of antimicrobial chemokines CXCL9 (net charge of +20) and CXCL10 (net charge +11)
to the cell wall of S. aureus likely via the positively charged patches on these protein surfaces [237].
Under certain situations, the antimicrobial peptide is generated by further processing of a precursor
protein. For example, antimicrobial thrombocidin-1 (TC-1) is produced by truncating two residues
from the C-terminus of the parent protein NAP-2 (i.e., neutrophil-activating peptide-2), which is
poorly active. NMR analysis revealed that the C-terminus of TC-1 is mobile. In contrast, the
C-terminus of NAP-2 is less mobile. It was proposed that the additional two residues locked the
C-terminus via electrostatic interactions [238]. The additional Asp residue could have masked the
positively-charged surface of the C-terminal helix that targets bacterial membranes. Likewise, insertion
of an acidic Glu to the N-terminal region of GF-17, a peptide corresponding to the major antimicrobial
region of human cathelicidin LL-37 [179], substantially reduced the peptide activity [239].
RNases. The structures of RNase 3 (Figure 4, panels A and B) and RNase 5 (panels C and D) were
solved by both X-ray diffraction and multi-dimensional NMR spectroscopy. Although RNase 3 is
dimeric in the crystal, the protein fold determined by the two techniques is similar. In addition, NMR
studies revealed two conformations for His114 of RNase 5 in solution [240]. The structures of RNase
2 and RNase 7 are given in Figure 4 (panels E and F). Unlike chemokines discussed above, the
antimicrobial region has been mapped to the N-terminus of RNase 7 [130,241]. In particular, a cluster
of lysines were identified as key elements for antibacterial activity (bold in Table 1). It is proposed
recently that the N-terminal antimicrobial function is conserved in the ribonuclease family [242]. One
may wonder why a protein is created for bacterial defense if only part of the chain is required to kill
pathogens. One possibility is the stability gain as part of the protein. Another possibility is that a
folded protein structure allows for the incorporation of a variety of active sites on the protein surface.
In certain cases, such functional sites may be overlapping [243]. In the case of RNase 7, the adjacent
active site and antimicrobial residues allows us to propose a yet-to-be-proved ―peel-and-kill‖ model. In
other words, binding to bacteria by the cationic amino acids is followed by digestion of pathogenic
nucleic acids. The multiple active sites also enable functional regulation. For example, an endogenous
molecule can bind to RNase 7 and regulates its antimicrobial activity [244]. This could be one of the
unique features of antimicrobial proteins distinct from small antimicrobial peptides.
Antimicrobial lectin RegIIIα. RegIIIα (or HIP/PAP) is a C-type lectin that binds peptidoglycan
carbohydrates of Gram-positive bacterial cell walls. The structural basis of this binding has been
elucidated (Figure 2D) [245]. Different from other C-type calcium-dependent lectins, the binding of
RegIIIα to peptidoglycans is calcium independent (i.e., lacking calcium-binding motif). The binding,
however, requires the ―EPN‖ motif and depends on sugar chain length. However, it seems that this
peptidoglycan binding serves as an early recognition step for the peptide action as it can create a pore
on bacterial membranes. The structure of the oligomeric form of the protein has recently been
determined by combining X-ray structure and electron microscopy data, providing insight into the
lethal step of bacterial killing by this intestine lectin [246].
Figure 4. 3D structures of human ribonucleases with antimicrobial activity. Shown are (A)
RNase 3 (dimeric crystal structure, PDB ID: 4A2O); (B) RNase 3 (NMR structure, PDB
ID: 2KB5); (C) RNase 5 (crystal structure, PDB ID: 1B1I); (D) RNase 5 (NMR structure,
PDB ID: 1AWZ); (E) RNase 2 (crystal structure, PDB ID: 2BZZ); and (F) RNase 7 (NMR
structure, PDB ID: 2HKY).
5. Mechanism of Action of Human Antimicrobial Peptides
It is generally believed that cationic AMPs target anionic bacterial membranes. In the past years,
significant advances have been made in elucidating the molecular targets of human AMPs. As
described below, human AMPs can interact with a variety of molecular targets either on the cell
surface (including membranes) or within the cells.
5.1. Targeting Bacterial Cell Wall
The molecular targets of AMPs are not limited to bacterial membranes. The assembly of HD-6 on
bacterial surface entangles bacteria, providing a new defense mechanism for human innate
immunity [123]. HNP-1 targets lipid II to block the biosynthesis of bacterial cell walls [247].
Böhling et al. found a close correlation of the hBD-3 activity with cell wall components [248]. A
subsequent study corroborated the binding of hBD-3 to lipid II as well [249]. In the APD [7,8], there
are 18 AMPs that use this mechanism to combat bacteria. Examples are nisins, mersacidin from
bacteria, plectasin from fungi, Cg-Def, an oyster defensin [250–252]. Hence, blocking bacterial cell
wall synthesis is a widely deployed innate defense mechanism. It is possible to identify small molecule
mimetics that bind bacterial cell walls [253,254]. Although not discussed here, other defensins can
directly recognize specific lipids in fungal membranes [255–257].
RegIII proteins are a family of lectins that can specifically recognize the carbohydrate portion of
bacteria. While RegIIIα targets the peptidoglycan carbohydrate backbone for Gram-positive bacterial
killing [245], mouse RegIIIβ associates with the lipid A portion of Gram-negative bacterial LPS [258].
In the case of murine RegIIIβ, amino acid residues in two structural motifs termed ―loop 1‖ and ―loop 2‖
are important for peptidoglycan and lipid A binding (Arg-135, Asp-142) and for the bactericidal
activity (Glu-134, Asn-136, and Asp-142).
It is well known that human lysozyme not only binds a single peptidoglycan chain but also cuts the
sugar repeats, thereby inhibiting bacterial cell wall synthesis [259]. In addition, some AMPs can
associate with cell surface proteins to interfere with the docking and entry of viruses such as human
immunodeficiency virus type 1 (HIV-1). For example, the association of SLPI with human annexin II
can avoid the attachment of viral lipid phosphatidylserine (PS) to the same protein in human
macrophages [260].
5.2. Targeting Bacterial Inner Membranes
Human cathelicidin LL-37 is a representative member in the helical family. It is proposed that
LL-37 disrupts bacterial membranes. The membrane disruption of LL-37 involves at least three steps.
First, the cationic peptide can recognize and coat the anionic surface of bacteria. With a classic
amphipathic helical structure, this cationic peptide prefers to target anionic bacterial membranes.
Second, LL-37 binds to the outer membranes and cross the outer membrane. Third, the peptide reaches
the inner membrane. It initially binds to the inner membrane parallel to the surface, which is the basis
for the carpet model [261]. At elevated concentrations, the peptide may disrupt the membranes by
micellization. Alternatively, the peptide might take a vertical position to form a pore [262].
The ability of hepcidin 25 (hep-25) and its isoform hepcidin 20 (hep-20) to perturb bacterial
membranes is markedly pH-dependent. The membrane disruption is more evident at acidic pH than at
neutral pH. At acidic conditions, histidines become positively charged and more effective in
membrane disruption [263].
While there is no agreement in the case of human LL-37 regarding the carpet or pore formation,
recent structural determination of dermcidin provides evidence for possible pore formation in bacteria
membranes. In the crystal structure [216], the peptide forms a hexamer, where two trimers are
connected by zinc (Figure 1C). It is proposed that this structure might be directly inserted into bacterial
membranes, serving as an ion channel.
In addition to α-defensins (HD-5 and HD-6), RegIII peptides are expressed to control the
microbiota and keep the bacteria away from the epithelial surface of intestine. In particular, specific
bacteria (e.g., Bifidobacterium breve NCC2950) can effectively induce the expression of RegIIIγ (an
ortholog of human RegIIIα) in the intestine of mice via the MyD88-Ticam1 pathway [264]. RegIIIα is
a lectin that binds to peptidoglycans of Gram-positive bacteria. However, it adopts a hexametic
membrane-permeating pore structure to kill bacteria [246]. Such a pore is reminiscent of other pore
structures solved for toxins [265–267]. The structure also provides a basis for selective killing of
Gram-positive bacteria such as L. monocytogenes, L. innocua, and E. faecalis but not Gram-negative
bacteria. This is because LPS, the major component of the outer membranes of Gram-negative
bacteria, inhibits the pore-forming activity of RegIIIα [246].
5.3. Cell-Penetrating Peptides and Intracellular Targets
There are other AMPs that may work primarily by binding DNA. Buforin is such an example [83].
This is not surprising because this AMP was derived from DNA-binding histone 2A. In addition, SLPI,
a small protein that inhibits elastase and cathepsin G, displayed antibacterial activity against
E. coli by binding nucleic acids [268]. It is also likely that AMPs kill bacteria by more than one
mechanism. For example, human LL-37 may first damage bacterial membranes followed by DNA
binding, leading to the shutdown of bacterial machinery [83].
Unlike human LL-37, histatin 5 caused only small membrane damaging effects [269]. To interpret
the killing effect of this peptide, two models have been proposed. In the first model, treatment of
C. albicans with histatin 5 induces the efflux of ATP and increases cell permeability to small
molecules, leading to ion imbalance. Thus, C. albicans cells respond by activating the osmotic stress
responding pathways to minimize ion loss. This model is supported by the fact that knocking out the
TRK1 gene that encodes a major K+ uptake system made histatin 5 ineffective. A second model was
also proposed based on the observation that the candida killing ability of histatin 5 is lost using a
mitochondrial respiration mutant or after treatment with sodium azide that inhibits cellular
metabolism [270]. In addition to the requirement of the mitochondrial respiration machinery [271],
cellular internalization of histatins is also facilitated by peptide binding to heat shock protein Ssa2p on
the surface of C. albicans [272]. This interference with mitochondrial respiration chain may
be responsible for the formation of reactive oxygen species (ROS), leading to cell death [273].
Vylkova et al. performed a DNA microarray study of the effect of histatin 5 and found that these two
models can be unified. This is because the oxidative stress could be produced as a secondary effect of
osmotic stress. This proposal is in line with the observation that the killing of histatin 5 is facilitated in
the presence of an osmotic agent sorbital but not an oxidant agent H2O2 [274]. It should be mentioned
that human GAPDH(2–32), an antifungal peptide derived from the highly conserved protein GAPDH,
can also enter C. albicans to induce apoptosis [275].
As a different mechanism to combat bacterial infection, some AMPs are reported to penetrate
immune cells and activate them to boost immune response. For example, chromagranin A-derived
peptides can penetrate neutrophils, bind to cytoplasmic calmodulin, and induce Ca2+ influx, leading to
neutrophil activation and immune system augmentation [276].
Granulysin is an effector molecule in the cytotoxic granules of cytotoxic T lymphocytes and natural
killer (NK) cells. It can kill intracellular pathogens in infected cells in the presence of perforin and to
induce a cytotoxic effect against tumor cells. Although perforin and granulysin can colocalize [277],
it is unclear how they work together in bacterial killing. A recent study reveals that perforin can form
pores that preferentially allow the entry of cationic molecules [278]. Thus, granulysin might have
entered the cell via the perforin pores, thereby providing yet another mechanism for intracellular
bacterial killing by forming a molecular pair.
6. Concluding Remarks and Potential Therapeutic Strategies
Human antimicrobial peptides and proteins occupy an important niche in the current research on
human host defense and innate immunity [1–6,279]. Except for antimicrobial protein lysozyme, which
was found in 1922, most of short cationic peptides were discovered after 1980 (Table 1). By the time
this article was written, over 100 human AMPs have been identified and characterized. They were
either isolated from human tissues or predicted from the human genome by bioinformatics. Although
genomic prediction constitutes an invaluable method, isolation from natural sources remains important
in determining the exact mature form of AMPs. The discovery story of LL-37 nicely illustrates this
point (Section 2.3). These peptides have diverse amino acid sequences (Table 1) and physical properties
(Table 2), leading to a panel of defense molecules with varying activities (Table 1). While psoriasin
and KDAMP primarily inhibit the growth of Gram-negative bacteria, RegIIIα is mainly active against
Gram-positive bacteria. In addition, histatins and drosomycin-like defensin are primarily fungicidal.
Many human AMPs such as LL-37 and defensins are broad-spectrum peptides against pathogens.
Remarkably, human AMPs are able to hinder bacterial growth by interactions with different targets,
ranging from surface molecules (e.g., cell walls), inner membranes, to intracellular molecules
(Table 3). Some AMPs can interact with two or more molecules. For example, the binding of RegIIIα
to peptidoglycans constitutes only the initial recognition step and subsequent pore formation in
bacterial membranes could be the lethal step [246].
Table 3. Select human antimicrobial peptides and their proposed targets.

APD ID AMP Structure Molecular target
181 HD-6 β Aggregate on bacterial surface
283 hBD-3 αβ Bacterial cell wall (lipid II)
176 HNP-1 β Bacterial cell wall (lipid II)
2257 Lysozyme α Cell wall carbohydrate
2071 RegIIIα αβ Membrane pores
310 LL-37 α Bacterial membranes and/or DNA
433 Dermcidin α Membranes ion channel
2017 hGAPDH(2-32) Unknown Intracellular targets of fungi
505 Histatin 5 α Intracellular mitochondria
Chromagranin Cytoplasmic calmodulin of
2352 Unknown
A-derived peptides neutrophils
Perforin generates a pore to allow
1161 Granulysin α granulysin to enter the cell and kill
intracellular bacteria
For interactions with different molecules, human AMPs are capable of adopting a variety of 3D
structures (Figures 1–4). It is clearly important to determine the structure to high quality so that the
molecular basis of these interactions can be uncovered accurately (reviewed in ref. [52]). It is also
important to correlate the structure with the active state of the peptide. In the case of dermcidin,
which oligomerizes on bacterial surface, the helix-bundle structure determined by X-ray
crystallography [216] should be more relevant. Likewise, the disulfide-bonded structure of HBD-1
does not explain peptide activity under reduced conditions [226]. Therefore, human AMPs are diverse
in terms of amino acid sequence, 3D structure, activity, and mechanism of action.
Many human AMPs are currently under close examination for their functional roles as well as
potential applications in detection and diagnosis of human diseases. The type and expression level of
human AMPs, if accurately mapped, may have clinical relevance. A clear variation in the expression level
of AMPs can serve as biomarkers for human diseases, such as eczema severity and cancer [280–283]. In
addition, this remarkable array of molecules may be used for detection, imaging, and diagnosis of
bacterial infection. An example of this application is based on the preferential association of
Technetium-99m labeled ubiquicidin with bacteria, enabling the physician to differentiate infection
from aseptic loosening of hip prostheses in 30 min with high accuracy [284–286].
The collection of human AMPs discussed herein also inspires us in developing novel therapeutics [287].
First, new antimicrobials may be developed using human AMPs as templates. The rationale is that
AMPs have remained potent for millions of years and are thus less prone to microbial resistance [1–6].
In particular, peptides with different structural scaffolds may kill the same bacterium by different
mechanisms (Table 3). Furthermore, the same peptide sequence can be tailored into various peptides
that selectively target pathogens such as Gram-positive, Gram-negative bacteria, or viruses [83]. This
is highly desirable for selective bacterial elimination without destroying the probiotic microbial flora.
The success of this line depends on whether a selected template will achieve the desired potency in
vivo against a target pathogen, low cytotoxicity to humans, stability to proteases, and cost-effective
production [287]. One can also consider alternative peptide forms. For instance, a pro-drug can be used
to reduce the cytotoxicity of AMPs if a mechanism can be found to release it when needed [288].
While portions of antimicrobial proteins are preferred to design novel antimicrobials [222,223,242,289],
a whole protein may also be considered. Unlike short peptides, the folded structure of proteins confers
stability to the action of proteases. However, the production of such a long polypeptide chain may
require recombinant expression in bacteria or cell-free systems [199].
Second, new strategies are actively sought to bring the invading pathogens under control. It is
appreciated that different receptors and signal pathways are activated in response to the invasion of
different microbes [65]. Of outstanding interest is that non-pathogen factors can also induce AMP
expression (Table 4). One of the earliest examples might be the light therapy invented by Niel
Finsen [290]. The establishment of a link between light therapy, vitamin D and human cathelicidin
LL-37 expression provides a completely different way for infection treatment. Instead of treating
patients with traditional antibiotics, doctors may be able to use light or vitamin D [291,292]. Indeed
using narrow-band UV B light, the level of vitamin D was increased in psoriasis patients (psoriasis is a
common autoimmune disease on skin) [293]. In addition, other small molecules such as butyrate can
induce LL-37 expression [294]. Components from Traditional Chinese Medicine may regulate the
AMP expression as well [295]. These factors may induce the expression of a single peptide or multiple
AMPs [296]. It is also possible that certain factors can work together to induce AMP expression.
While cyclic AMP and butyrate synergistically stimulate the expression of chicken β-defensin 9 [297],
4-phenylbutyrate (PBA) and 1,25-dihydroxyvitamin D3 (or lactose) can induce AMP gene expression
synergistically [294,298]. It appears that stimulation of LL-37 expression by histone deacetylase
(HDAC) inhibitors is cell dependent. Trichostatin and sodium butyrate increased the peptide
expression in human NCI-H292 airway epithelial cells but not in the primary cultures of normal nasal
epithelial cells [299]. However, the induction of the human LL-37 expression may not be
a general approach for bacterial clearance. During Salmonella enterica infection of human
monocyte-derived macrophages, LL-37 is neither induced nor required for bacterial clearance [300].
Table 4. Some known factors that induce antimicrobial peptide expression

Factor AMP induced Cells Ref
Bacteria/LPS LL-37, HBD-2 keratinocytes [296]
TNF-α LL-37, HBD-2 keratinocytes [296]
UV Light LL-37, HBD-2, chemerin keratinocytes [296,301]
neutrophil progenitors and
Vitamin D3 LL-37 [302,303]
EBV-transformed B cells
colonic epithelial cells
Lactose LL-37 T84, THP-1 monocytes [304]
and macrophages
LL-37; human HT-29 colonic
Short-chain fatty acids pBD-2, pBD-3, pEP2C, and epithelial cells and U-937 [305,306]
protegrins monocytic cells;
human colon cells, HCT-
hBD-1; 116;
Isoleucine [307–309]
epithelial defensins bovine kidney epithelial
cells
Arginine hBD-1 [307]
116
human keratinocyte
Ca2+ hBD-2, hBD-3 [310]
monolayers
LL-37; Caco-2 cell;
Zn2+ [311]
pBD-1, pBD-2, pBD-3 Intestinal epithelial cells
colon, gastric and
Butyrate LL-37 [312]
hepatocellular cells
Albumin hBD-1 [307]
116
macrophages and primary
Cyclic AMP/Butyrate Chicken β-defensin 9 [297]
jejunal explants
immortalized human
Phenylbutyrate/1,25-
cathelicidins bronchial epithelial cell [298]
dihydroxyvitamin D3
line VA10
Finally, immune modulation peptides may find therapeutic use because they do not act on microbes
directly and thereby are less likely to induce antimicrobial resistance [4,5]. Immune modulation is
activated via peptide binding to host cell receptors that initiate various signal transduction pathways.
Recently, a natural peptide was found to have immune modulating activity but no antimicrobial
activity [313]. Besides engineering peptides with distinct properties, there has been growing interest in
elucidating the bacterial mechanisms in generating resistance to AMPs or by subverting host immune
systems [314,315]. It can be anticipated that new therapeutic approaches will continue to emerge from
our understanding of the host-pathogen interactions. All these strategies will facilitate the development
of AMPs into novel antimicrobials to meet the challenge of antibiotics-resistant superbugs, RNA viral
infections and difficult-to-treat cancers [287].
Acknowledgements
The author is grateful for the support of the fundings from the NIH (R56AI081975) and the state of
Nebraska during this study.
Conflicts of Interest
The author does not declare a conflict of interest.
References
1. Boman, H.G. Antibacterial peptides: Basic facts and emerging concepts. J. Inter. Med. 2003,
254, 197–215.
2. Ganz, T.; Lehrer, R.I. Defensins. Curr. Opin. Immunol. 1994, 6, 584–589.
3. Zasloff, M. Antimicrobial peptides of multicellullar organisms. Nature 2002, 415, 359–365.
4. Hancock, R.E.W.; Sahl, H.G. Antimicrobial and host-defense peptides as new anti-infective
therapeutic strategies. Nat. Biotechnol. 2006, 24, 1551–1557.
5. Lai, Y.; Gallo, R.L. AMPed up immunity: How antimicrobial peptides have multiple roles in
immune defense. Trends Immunol. 2009, 30, 131–141.
6. Yount, N.Y.; Yeaman, M.R. Emerging themes and therapeutic prospects for anti-infective
peptides. Annu. Rev. Pharmacol. Toxicol. 2012, 52, 337–360.
7. Wang, Z.; Wang, G. APD: The antimicrobial peptide database. Nucleic Acids Res. 2004, 32,
D590–D592.
8. Wang, G.; Li, X.; Wang, Z. The updated antimicrobial peptide database and its application in
peptide design. Nucleic Acids Res. 2009, 37, D933–D937.
9. Marchini, G.; Lindow, S.; Brismar, H.; Ståbi, B.; Berggren, V.; Ulfgren, A.K.; Lonne-Rahm, S.;
Agerberth, B.; Gudmundsson, G.H. The newborn infant is protected by an innate antimicrobial
barrier: Peptide antibiotics are present in the skin and vernix caseosa. Br. J. Dermatol. 2002, 147,
1127–1134.
10. Gschwandtner, M.; Zhong, S.; Tschachler, A.; Mlitz, V.; Karner, S.; Elbe-Bürger, A.;
Mildner, M. Fetal Human Keratinocytes Produce Large Amounts of Antimicrobial Peptides:
Involvement of Histone-Methylation Processes. J. Invest. Dermatol. 2014, doi:10.1038/jid.2014.165.
11. Wittersheim, M.; Cordes, J.; Meyer-Hoffert, U.; Harder, J.; Hedderich, J.; Gläser, R. Differential
expression and in vivo secretion of the antimicrobial peptides psoriasin (S100A7), RNase 7,
human beta-defensin-2 and -3 in healthy human skin. Exp. Dermatol. 2013, 22, 364–366.
12. Gläser, R.; Meyer-Hoffert, U.; Harder, J.; Cordes, J.; Wittersheim, M.; Kobliakova, J.;
Fölster-Holst, R.; Proksch, E.; Schröder, J.M.; Schwarz, T. The antimicrobial protein psoriasin
(S100A7) is upregulated in atopic dermatitis and after experimental skin barrier disruption.
J. Invest. Dermatol. 2009, 129, 641–649.
13. McDermott, A.M. Antimicrobial compounds in tears. Exp. Eye Res. 2013, 117, 53–61.
14. Underwood, M.; Bakaletz, L. Innate immunity and the role of defensins in otitis media. Curr.
Allergy Asthma Rep. 2011, 11, 499–507.
15. Sato, J.; Nishimura, M.; Yamazaki, M.; Yoshida, K.; Kurashige, Y.; Saitoh, M.; Abiko, Y.
Expression profile of drosomycin-like defensin in oral epithelium and oral carcinoma cell lines.
Arch. Oral Biol. 2013, 58, 279–285.
16. Da Silva, B.R.; de Freitas, V.A.; Nascimento-Neto, L.G.; Carneiro, V.A.; Arruda, F.V.;
de Aguiar, A.S.; Cavada, B.S.; Teixeira, E.H. Antimicrobial peptide control of pathogenic
microorganisms of the oral cavity: A review of the literature. Peptides 2012, 36, 315–321.
17. Tollner, T.L.; Bevins, C.L.; Cherr, G.N. Multifunctional glycoprotein DEFB126—A curious
story of defensin-clad spermatozoa. Nat. Rev. Urol. 2012, 9, 365–375.
18. Yu, H.; Dong, J.; Gu, Y.; Liu, H.; Xin, A.; Shi, H.; Sun, F.; Zhang, Y.; Lin, D.; Diao, H. The
novel human β-defensin 114 regulates lipopolysaccharide (LPS)-mediated inflammation and
protects sperm from motility loss. J. Biol. Chem. 2013, 288, 12270–12282.
19. Tollner, T.L.; Yudin, A.I.; Tarantal, A.F.; Treece, C.A.; Overstreet, J.W.; Cherr, G.N.
Beta-defensin 126 on the surface of macaque sperm mediates attachment of sperm to oviductal
epithelia. Biol. Reprod. 2008, 78, 400–412.
20. Fleming, A. On a remarkable bacteriolytic element found in tissues and secretions. Proc. R. Soc. B
1922, 93, 306–317.
21. Selsted, M.E.; Harwig, S.S.; Ganz, T.; Schilling, J.W.; Lehrer, R.I. Primary structures of three
human neutrophil defensins. J. Clin. Invest. 1985, 76, 1436–1439.
22. Oppenheim, F.G.; Xu, T.; McMillian, F.M.; Levitz, S.M.; Diamond, R.D.; Offner, G.D.;
Troxler, R.F. Histatins, a novel family of histidine-rich proteins in human parotid secretion.
Isolation, characterization, primary structure, and fungistatic effects on Candida albicans.
J. Biol. Chem. 1988, 263, 7472–7477.
23. Wilde, C.G.; Griffith, J.E.; Marra, M.N.; Snable, J.L.; Scott, R.W. Purification and
characterization of human neutrophil peptide 4, a novel member of the defensin family. J. Biol.
Chem. 1989, 264, 11200–11203.
24. Hamann, K.J.; Gleich, G.J.; Checkel, J.L.; Loegering, D.A.; McCall, J.W.; Barker, R.L. In vitro
killing of microfilariae of Brugia pahangi and Brugia malayi by eosinophil granule proteins.
J Immunol. 1990, 144, 3166–3173.
25. Jones, D.E.; Bevins, C.L. Paneth cells of the human small intestine express an antimicrobial
peptide gene. J. Biol. Chem. 1992, 267, 23216–23225.
26. Jones, D.E.; Bevins, C.L. Defensin-6 mRNA in human Paneth cells: Implications for
antimicrobial peptides in host defense of the human bowel. FEBS Lett. 1993, 315, 187–192.
27. Bensch, K.W.; Raida, M.; Mägert, H.J.; Schulz-Knappe, P.; Forssmann, W.G. hBD-1: A novel
beta-defensin from human plasma. FEBS Lett. 1995, 368, 331–335.
28. Agerberth, B.; Gunne, H.; Odeberg, J.; Kogner, P.; Boman, H.G.; Gudmundsson, G.H. FALL-39,
a putative human peptide antibiotic, is cysteine-free and expressed in bone marrow and testis,
Proc. Natl. Acad. Sci. USA 1995, 92, 195–199.
29. Larrick, J.W.; Hirata, M.; Balint, R.F.; Lee, J.; Zhong, J.; Wright, S.C. Human CAP18: A novel
antimicrobial lipopolysaccharide-binding protein. Infect. Immun. 1995, 63, 1291–1297.
30. Cowland, J.B.; Johnsen, A.H.; Borregaard, N. hCAP-18, a cathelin/pro-bactenecin-like protein of
human neutrophil specific granules. FEBS Lett. 1995, 368, 173–176.
31. Harder, J.; Bartels, J.; Christophers, E.; Schröder, J.M. A peptide antibiotic from human skin.
Nature 1997, 387, 861.
32. Stenger, S.; Hanson, D.A.; Teitelbaum, R.; Dewan, P.; Niazi, K.R.; Froelich, C.J.; Ganz, T.;
Thoma-Uszynski, S.; Melián, A.; Bogdan, C.; et al. An antimicrobial activity of cytolytic T cells
mediated by granulysin. Science 1998, 282, 121–125.
33. Hieshima, K.; Ohtani, H.; Shibano, M.; Izawa, D.; Nakayama, T.; Kawasaki, Y.; Shiba, F.;
Shiota, M.; Katou, F.; Saito, T.; et al. CCL28 has dual roles in mucosal immunity as a chemokine
with broad-spectrum antimicrobial activity. J. Immunol. 2003, 170, 1452–1461
34. Krijgsveld, J.; Zaat, S.A.; Meeldijk, J.; van Veelen, P.A.; Fang, G.; Poolman, B.; Brandt, E.;
Ehlert, J.E.; Kuijpers, A.J.; Engbers, G.H.; et al. Thrombocidins, microbicidal proteins from
human blood platelets, are C-terminal deletion products of CXC chemokines. J. Biol. Chem.
2000, 275, 20374–20381.
35. Krause, A.; Neitz, S.; Mägert, H.J.; Schulz, A.; Forssmann, W.G.; Schulz-Knappe, P.;
Adermann, K. LEAP-1, a novel highly disulfide-bonded human peptide, exhibits antimicrobial
activity. FEBS Lett. 2000, 480, 147–150.
36. Cutuli, M.; Cristiani, S.; Lipton, J.M.; Catania, A. Antimicrobial effects of alpha-MSH peptides.
J. Leukoc. Biol. 2000, 67, 233–239.
37. Harder, J.; Bartels, J.; Christophers, E.; Schroeder, J.M. Isolation and characterization of human
deta-defensin-3, a novel human inducible peptide antibiotic. J. Biol. Chem. 2001, 276,
5707–5713.
38. García, J.R.; Krause, A.; Schulz, S.; Rodríguez-Jiménez, F.J.; Klüver, E.; Adermann, K.;
Forssmann, U.; Frimpong-Boateng, A.; Bals, R.; Forssmann, W.G. Human beta-defensin 4: A
novel inducible peptide with a specific salt-sensitive spectrum of antimicrobial activity. FASEB J.
2001, 15, 1819–1821.
39. Schittek, B.; Hipfel, R.; Sauer, B.; Bauer, J.; Kalbacher, H.; Stevanovic, S.; Schirle, M.;
Schroeder, K.; Blin, N.; Meier, F.; et al. Dermcidin: A novel human antibiotic peptide secreted
by sweat glands. Nat. Immunol. 2001, 2, 1133–1137.
40. Harder, J.; Schroder, J.M. RNase 7, a novel innate immune defense antimicrobial protein of
healthy human skin. J. Biol. Chem. 2002, 277, 46779–46784.
41. Hooper, L.V.; Stappenbeck, T.S.; Hong, C.V.; Gordon, J.I. Angiogenins: A new class of
microbicidal proteins involved in innate immunity. Nat. Immunol. 2003, 4, 269–273.
42. Yang, D.; Chen, Q.; Hoover, D.M.; Staley, P.; Tucker, K.D.; Lubkowski, J.; Oppenheim, J.J.
Many chemokines including CCL20/MIP-3alpha display antimicrobial activity. J. Leukoc. Biol.
2003, 74, 448–455.
43. Gläser, R.; Harder, J.; Lange, H.; Bartels, J.; Christophers, E.; Schröder, J.M. Antimicrobial
psoriasin (S100A7) protects human skin from Escherichia coli infection. Nat. Immunol. 2005, 6,
57–64.
44. Cash, H.L.; Whitham, C.V.; Behrendt, C.L.; Hooper, L.V. Symbiotic bacteria direct expression
of an intestinal bactericidal lectin. Science 2006, 313, 1126–1130.
45. El Karim, I.A.; Linden, G.J.; Orr, D.F.; Lundy, F.T. Antimicrobial activity of neuropeptides
against a range of micro-organisms from skin, oral, respiratory and gastrointestinal tract sites.
J. Neuroimmunol. 2008, 200, 11–16.
46. Simon, A.; Kullberg, B.J.; Tripet, B.; Boerman, O.C.; Zeeuwen, P.; van der Ven-Jongekrijg, J.;
Verweij, P.; Schalkwijk, J.; Hodges, R.; van der Meer, J.W.; et al. Drosomycin-like defensin, a
human homologue of Drosophila melanogaster drosomycin with antifungal activity. Antimicrob.
Agents Chemother. 2008, 52, 1407–1412.
47. Marischen, L.; Wesch, D.; Schröder, J.M.; Wiedow, O.; Kabelitz, D. Human γδ T cells produce
the protease inhibitor and antimicrobial peptide elafin. Scand. J. Immunol. 2009, 70, 547–552.
48. Soscia, S.J.; Kirby, J.E.; Washicosky, K.J.; Tucker, S.M.; Ingelsson, M.; Hyman, B.;
Burton, M.A.; Goldstein, L.E.; Duong, S.; Tanzi, R.E.; et al. The Alzheimer’s disease-associated
amyloid beta-protein is an antimicrobial peptide. PLoS One 2010, 5, e9505.
49. Kulig, P.; Kantyka, T.; Zabel, B.A.; Banas, M.; Chyra, A.; Stefanska, A.; Tu, H.; Allen, S.J.;
Handel, T.M.; Kozik, A.; et al. Regulation of chemerin chemoattractant and antibacterial activity
by human cysteine cathepsins. J. Immunol. 2011, 187, 1403–1410.
50. Wang, L.; Liu, Q.; Chen, J.C.; Cui, Y.X.; Zhou, B.; Chen, Y.X.; Zhao, Y.F.; Li, Y.M.
Antimicrobial activity of human islet amyloid polypeptides: An insight into amyloid peptides’
connection with antimicrobial peptides. Biol. Chem. 2012, 393, 641–646.
51. Tam, C.; Mun, J.J.; Evans, D.J.; Fleiszig, S.M. Cytokeratins mediate epithelial innate defense
through their antimicrobial properties. J. Clin. Invest. 2012, 122, 3665–3677.
52. Wang, G. Database-guided discovery of potent peptides to combat HIV-1 or superbugs.
Pharmaceuticals 2013, 6, 728–758.
53. Zhao, C.; Wang, I.; Lehrer, R.I. Widespread expression of beta-defensin hBD-1 in human
secretory glands and epithelial cells. FEBS Lett. 1996, 396, 319–322.
54. Ouellette, A.J.; Greco, R.M.; James, M.; Frederick, D.; Naftilan, J.; Fallon, J.T. Developmental
regulation of cryptdin, a corticostatin/defensin precursor mRNA in mouse small intestinal crypt
epithelium. J. Cell Biol. 1989, 108, 1687–1695.
55. Ganz, T. Defensins in the urinary tract and other tissues. J. Infect. Dis. 2001, 183 Suppl 1,
S41–S42.
56. Valore, E.V.; Park, C.H.; Quayle, A.J.; Wiles, K.R.; McCray PB, Jr.; Ganz, T. Human
beta-defensin-1: An antimicrobial peptide of urogenital tissues. J. Clin. Invest. 1998, 101,
1633–1642.
57. Goldman, M.J.; Anderson, G.M.; Stolzenberg, E.D.; Kari, U.P.; Zasloff, M.; Wilson, J.M.
Human beta-defensin-1 is a salt-sensitive antibiotic in lung that is inactivated in cystic fibrosis.
Cell 1997, 88, 553–560.
58. Bals, R.; Wang, X.; Wu, Z.; Freeman, T.; Bafna, V.; Zasloff, M.; Wilson, J.M. Human
beta-defensin 2 is a salt-sensitive peptide antibiotic expressed in human lung. J. Clin. Invest.
1998, 102, 874–880.
59. Jia, H.P.; Schutte, B.C.; Schudy, A.; Linzmeier, R.; Guthmiller, J.M.; Johnson, G.K.; Tack, B.F.;
Mitros, J.P.; Rosenthal, A.; Ganz, T.; et al. Discovery of new human beta-defensins using a
genomics-based approach. Gene 2001, 263, 211–218.
60. García, J.R.; Jaumann, F.; Schulz, S.; Krause, A.; Rodríguez-Jiménez, J.; Forssmann, U.;
Adermann, K.; Klüver, E.; Vogelmeier, C.; Becker, D.; et al. Identification of a novel,
multifunctional beta-defensin (human beta-defensin 3) with specific antimicrobial activity. Its
interaction with plasma membranes of Xenopus oocytes and the induction of macrophage
chemoattraction. Cell Tissue Res. 2001, 306, 257–264.
61. Scheetz, T.; Bartlett, J.A.; Walters, J.D.; Schutte, B.C.; Casavant, T.L.; McCray, P.B, Jr.
Genomics-based approaches to gene discovery in innate immunity. Immunol. Rev. 2002, 190,
137–145.
62. Huang, L.; Leong, S.S.; Jiang, R. Soluble fusion expression and characterization of bioactive
human beta-defensin 26 and 27. Appl. Microbiol. Biotechnol. 2009, 84, 301–308.
63. Schulz, A.; Klüver, E.; Schulz-Maronde, S.; Adermann, K. Engineering disulfide bonds of the
novel human beta-defensins hBD-27 and hBD-28: Differences in disulfide formation and
biological activity among human beta-defensins. Biopolymers 2005, 80, 34–49.
64. Xin, A.; Zhao, Y.; Yu, H.; Shi, H.; Liu, H.; Diao, H.; Zhang, Y. Soluble fusion expression,
characterization and localization of human β-defensin 6. Mol. Med. Rep. 2014, 9, 149–155.
65. Lemaitre, B.; Reichhart, J.M.; Hoffmann, J.A. Drosophila host defense: Differential induction of
antimicrobial peptide genes after infection by various classes of microorganisms. Proc. Natl.
Acad. Sci. USA 1997, 94, 14614–14619.
66. Selsted, M.E. Theta-defensins: cyclic antimicrobial peptides produced by binary ligation of
truncated alpha-defensins. Curr Protein Pept Sci. 2004, 5, 365-371.
67. Tang, Y.Q.; Yuan, J.; Osapay, G.; Osapay, K.; Tran, D.; Miller, C.J.; Ouellette, A.J.;
Selsted, M.E. A cyclic antimicrobial peptide produced in primate leukocytes by the ligation of
two truncated alpha-defensins. Science 1999, 286, 498–502.
68. Cole, A.M.; Wang, W.; Waring, A.J.; Lehrer, R.I. Retrocyclins: Using past as prologue. Curr.
Protein Pept. Sci. 2004, 5, 373–381.
69. Yang, C.; Boone, L.; Nguyen, T.X.; Rudolph, D.; Limpakarnjanarat, K.; Mastro, T.D.;
Tappero, J.; Cole, A.M.; Lal, R.B. Theta-Defensin pseudogenes in HIV-1-exposed, persistently
seronegative female sex-workers from Thailand. Infect Genet Evol. 2005, 5, 11–15.
70. Lehrer, R.I.; Cole, A.M.; Selsted, M.E. θ-Defensins: Cyclic peptides with endless potential.
J. Biol. Chem. 2012, 287, 27014–27019
71. Troxler, R.F.; Offner, G.D.; Xu, T.; Vanderspek, J.C.; Oppenheim, F.G. Structural relationship
between human salivary histatins. J. Dent. Res. 1990, 69, 2–6.
72. Sabatini, L.M.; Azen, E.A. Histatins, a family of salivary histidine-rich proteins, are encoded by
at least two loci (HIS1 and HIS2). Biochem. Biophys. Res. Commun. 1989, 160, 495–502.
73. vanderSpek, J.C.; Wyandt, H.E.; Skare, J.C.; Milunsky, A.; Oppenheim, F.G.; Troxler, R.F.
Localization of the genes for histatins to human chromosome 4q13 and tissue distribution of the
mRNAs. Am. J. Hum. Genet. 1989, 45, 381–387.
74. Nizet, V.; Ohtake, T.; Lauth, X.; Trowbridge, J.; Rudisill, J.; Dorschner, R.A.; Pestonjamasp, V.;
Piraino, J.; Huttner, K.; Gallo, R.L. Innate antimicrobial peptide protects the skin from invasive
bacterial infection. Nature 2001, 414, 454–457.
75. Braff, M.H.; Zaiou, M.; Fierer, J.; Nizet, V.; Gallo, R.L. Keratinocyte production of cathelicidin
provides direct activity against bacterial skin pathogens. Infect. Immun. 2005, 73, 6771–6781.
76. Lee, P.H.; Ohtake, T.; Zaiou, M.; Murakami, M.; Rudisill, J.A.; Lin, K.H.; Gallo, R.L.
Expression of an additional cathelicidin antimicrobial peptide protects against bacterial skin
infection. Proc. Natl. Acad. Sci. USA 2005, 102, 3750–3755.
77. Romeo, D.; Skerlavaj, B.; Bolognesi, M.; Gennaro, R. Structure and bactericidal activity of an
antibiotic dodecapeptide purified from bovine neutrophils. J. Biol. Chem. 1988, 263, 9573–9575.
78. Zanetti, M. Cathelicidins, multifunctional peptides of the innate immunity. J. Leukoc. Biol. 2004,
75, 39–48.
79. Gudmundsson, G.H.; Agerberth, B.; Odeberg, J.; Bergman, T.; Olsson, B.; Salcedo, R. The
human gene FALL-39 and processing of the cathelin precursor to the antibacterial peptide LL-37
in granulocytes, Eur. J. Biochem. 1996, 238, 325–332.
80. Sørensen, O.E.; Gram, L.; Johnsen, A.H.; Andersson, E.; Bangsbøll, S.; Tjabringa, G.S.;
Hiemstra, P.S.; Malm, J.; Egesten, A.; Borregaard, N. Processing of seminal plasma hCAP-18 to
ALL-38 by gastricsin: A novel mechanism of generating antimicrobial peptides in vagina.
J. Biol. Chem. 2003, 278, 28540–28546.
81. Zhao, H.; Lee, W.H.; Shen, J.H.; Li, H.; Zhang, Y. Identification of novel semenogelin I-derived
antimicrobial peptide from liquefied human seminal plasma. Peptides 2008, 29, 505–511
82. Yamasaki, K.; Schauber, J.; Coda, A.; Lin, H.; Dorschner, R.A.; Schechter, N.M.; Bonnart, C.;
Descargues, P.; Hovnanian, A.; Gallo, R.L. Kallikrein-mediated proteolysis regulates the
antimicrobial effects of cathelicidins in skin. FASEB J. 2006, 20, 2068–2080.
83. Wang, G.; Mishra, B.; Epand, R.F.; Epand, R.M. High-quality 3D structures shine light on
antibacterial, anti-biofilm and antiviral activities of human cathelicidin LL-37 and its fragments.
Biochim. Biophys. Acta 2014, doi: 10.1016/j.bbamem.2014.01.016.
84. Tailor, R.H.; Acland, D.P.; Attenborough, S.; Cammue, B.P.; Evans, I.J.; Osborn, R.W.;
Ray, J.A.; Rees, S.B.; Broekaert, W.F. A novel family of small cysteine-rich antimicrobial
peptides from seed of Impatiens balsamina is derived from a single precursor protein. J. Biol.
Chem. 1997, 272, 24480–24487.
85. Scocchi, M.; Bontempo, D.; Boscolo, S.; Tomasinsig, L.; Giulotto, E.; Zanetti, M. Novel
cathelicidins in horse leukocytes. FEBS Lett. 1999, 457, 459–464.
86. Anderson, R.C.; Yu, P.L. Isolation and characterisation of proline/arginine-rich cathelicidin
peptides from ovine neutrophils. Biochem. Biophys. Res. Commun. 2003, 312, 1139–1146.
87. Skerlavaj, B.; Benincasa, M.; Risso, A.; Zanetti, M.; Gennaro, R. SMAP-29: A potent
antibacterial and antifungal peptide from sheep leukocytes. FEBS Lett. 1999, 463, 58–62.
88. Castiglioni, B.; Scocchi, M.; Zanetti, M.; Ferretti, L. Six antimicrobial peptide genes of the
cathelicidin family map to bovine chromosome 22q24 by fluorescence in situ hybridization.
Cytogenet. Cell Genet. 1996, 75, 240–242.
89. Murakami, M.; Ohtake, T.; Dorschner, R.A.; Schittek, B.; Garbe, C.; Gallo, R.L. Cathelicidin
anti-microbial peptide expression in sweat, an innate defense system for the skin. J. Invest.
Dermatol. 2002, 119, 1090–1095.
90. Rieg, S.; Garbe, C.; Sauer, B.; Kalbacher, H.; Schittek, B. Dermcidin is constitutively produced
by eccrine sweat glands and is not induced in epidermal cells under inflammatory skin
conditions. Br. J. Dermatol. 2004, 151, 534–539.
91. Rieg, S.; Saborowski, V.; Kern, W.V.; Jonas, D.; Bruckner-Tuderman, L.; Hofmann, S.C.
Expression of the sweat-derived innate defence antimicrobial peptide dermcidin is not impaired
in Staphylococcus aureus colonization or recurrent skin infections. Clin. Exp. Dermatol. 2013,
doi:10.1111/ced.12189. tam
92. Schittek, B. The multiple facets of dermcidin in cell survival and host defense. J. Innate Immun.
2012, 4, 349–360.
93. Ghosh, R.; Maji, U.K.; Bhattacharya, R.; Sinha, A.K. The role of dermcidin isoform 2: A
two-faceted atherosclerotic risk factor for coronary artery disease and the effect of acetyl
salicylic acid on it. Thrombosis 2012, 2012, 987932.
94. Park, C.H.; Valore, E.V.; Waring, A.J.; Ganz, T. Hepcidin, a urinary antimicrobial peptide
synthesized in the liver. J. Biol. Chem. 2001, 276, 7806–7801.
95. Jordan, J.B.; Poppe, L.; Haniu, M.; Arvedson, T.; Syed, R.; Li, V.; Kohno, H.; Kim, H.;
Schnier, P.D.; Harvey, T.S.; et al. Hepcidin revisited, disulfide connectivity, dynamics, and
structure. J. Biol. Chem. 2009, 284, 24155–24167.
96. Pigeon, C.; Ilyin, G.; Courselaud, B.; Leroyer, P.; Turlin, B.; Brissot, P.; Loréal, O. A new mouse
liver-specific gene, encoding a protein homologous to human antimicrobial peptide hepcidin, is
overexpressed during iron overload. J. Biol. Chem. 2001, 276, 7811–7819.
97. Roetto, A.; Papanikolaou, G.; Politou, M.; Alberti, F.; Girelli, D.; Christakis, J.; Loukopoulos, D.;
Camaschella, C. Mutant antimicrobial peptide hepcidin is associated with severe juvenile
hemochromatosis. Nat. Genet. 2003, 33, 21–22.
98. Krause, A.; Sillard, R.; Kleemeier, B.; Klüver, E.; Maronde, E.; Conejo-García, J.R.;
Forssmann, W.G.; Schulz-Knappe, P.; Nehls, M.C.; Wattler, F.; et al. Isolation and biochemical
characterization of LEAP-2, a novel blood peptide expressed in the liver. Protein Sci. 2003, 12,
143–152.
99. Park, C.B.; Kim, M.S.; Kim, S.C. A novel antimicrobial peptide from Bufo bufo gargarizans.
Biochem. Biophys. Res. Commun. 1996, 218, 408–413.
100. Minn, I.; Kim, H.S.; Kim, S.C. Antimicrobial peptides derived from pepsinogens in the stomach
of the bullfrog, Rana catesbeiana. Biochim. Biophys. Acta 1998, 1407, 31–39.
101. Cole, A.M.; Kim, Y.H.; Tahk, S.; Hong, T.; Weis, P.; Waring, A.J.; Ganz, T. Calcitermin, a
novel antimicrobial peptide isolated from human airway secretions. FEBS Lett. 2001, 504, 5–10.
102. Park, C.J.; Park, C.B.; Hong, S.S.; Lee, H.S.; Lee, S.Y.; Kim, S.C. Characterization and cDNA
cloning of two glycine- and histidine-rich antimicrobial peptides from the roots of shepherd’s
purse, Capsella. bursa-pastoris. Plant. Mol. Biol. 2000, 44, 187–197.
103. Chang, C.I.; Pleguezuelos, O.; Zhang, Y.A.; Zou, J.; Secombes, C.J. Identification of a novel
cathelicidin gene in the rainbow trout, Oncorhynchus. mykiss. Infect. Immun. 2005, 73,
5053–5064.
104. Bayer, A.; Freund, S.; Jung, G. Post-translational heterocyclic backbone modifications in the
43-peptide antibiotic microcin B17. Structure elucidation and NMR study of a 13C,15N-labelled
gyrase inhibitor. Eur. J. Biochem. 1995, 234, 414–426.
105. Sousa1, J.C.; Berto1, R.F.; Gois, E.A.; Fontenele-Cardi, N.C.; Honório-Júnior, J.E.; Konno, K.;
Richardson, M.; Rocha, M.F.; Camargo, A.A.; Pimenta, D.C.; et al. Leptoglycin: A new
Glycine/Leucine-rich antimicrobial peptide isolated from the skin secretion of the South
American frog Leptodactylus. pentadactylus (Leptodactylidae). Toxicon 2009, 54, 23–32.
106. Couillault, C.; Pujol, N.; Reboul, J.; Sabatier, L.; Guichou, J.F.; Kohara, Y.; Ewbank, J.J.
TLR-independent control of innate immunity in Caenorhabditis elegans by the TIR domain
adaptor protein TIR-1, an ortholog of human SARM. Nat. Immunol. 2004, 5, 488–494.
107. Hao, X.; Yang, H.; Wei, L.; Yang, S.; Zhu, W.; Ma, D.; Yu, H.; Lai, R. Amphibian cathelicidin
fills the evolutionary gap of cathelicidin in vertebrate. Amino Acids 2012, 43, 677–685.
108. Lorenzini, D.M.; da Silva, P.I., Jr.; Fogaça, A.C.; Bulet, P.; Daffre, S. Acanthoscurrin: A novel
glycine-rich antimicrobial peptide constitutively expressed in the hemocytes of the spider
Acanthoscurria. gomesiana. Dev. Comp. Immunol. 2003, 27, 781–791.
109. Zeng, Y. Procambarin: A glycine-rich peptide found in the haemocytes of red swamp crayfish
Procambarus clarkii and its response to white spot syndrome virus challenge. Fish. Shellfish
Immunol. 2013, 35, 407–412.
110. Lee, S.Y.; Moon, H.J.; Kurata, S.; Natori, S.; Lee, B.L. Purification and cDNA cloning of an
antifungal protein from the hemolymph of Holotrichia. diomphalia larvae. Biol. Pharm. Bull.
1995, 18, 1049–1052.
111. Hiemstra, P.S.; van den Barselaar, M.T.; Roest, M.; Nibbering, P.H.; van Furth, R. Ubiquicidin, a
novel murine microbicidal protein present in the cytosolic fraction of macrophages. J. Leukoc.
Biol. 1999, 66, 423–428.
112. Linde, C.M.; Grundström, S.; Nordling, E.; Refai, E.; Brennan, P.J.; Andersson, M. Conserved
structure and function in the granulysin and NK-lysin peptide family. Infect. Immun. 2005, 73,
6332–6339.
113. Drannik, A.G.; Nag, K.; Sallenave, J.M.; Rosenthal, K.L. Antiviral activity of trappin-2 and
elafin in vitro and in vivo against genital herpes. J. Virol. 2013, 87, 7526–7538.
114. Strub, J.M.; Garcia-Sablone, P.; Lonning, K.; Taupenot, L.; Hubert, P.; van Dorsselaer, A.;
Aunis, D.; Metz-Boutigue, M.H. Processing of chromogranin B in bovine adrenal medulla.
Identification of secretolytin, the endogenous C-terminal fragment of residues 614–626 with
antibacterial activity. Eur. J. Biochem. 1995, 229, 356–368.
115. Lugardon, K.; Raffner, R.; Goumon, Y.; Corti, A.; Delmas, A.; Bulet, P.; Aunis, D.;
Metz-Boutigue, M.H. Antibacterial and antifungal activities of vasostatin-1, the N-terminal
fragment of chromogranin A. J. Biol. Chem. 2000, 275, 10745–10753.
116. Briolat, J.; Wu, S.D.; Mahata, S.K.; Gonthier, B.; Bagnard, D.; Chasserot-Golaz, S.; Helle, K.B.;
Aunis, D.; Metz-Boutigue, M.H. New antimicrobial activity for the catecholamine
release-inhibitory peptide from chromogranin A. Cell. Mol. Life Sci. 2005, 62, 377–385.
117. Goumon, Y.; Strub, J.M.; Moniatte, M.; Nullans, G.; Poteur, L.; Hubert, P.; van Dorsselaer, A.;
Aunis, D.; Metz-Boutigue, M.H. The C-terminal bisphosphorylated proenkephalin-A-(209-237)-
peptide from adrenal medullary chromaffin granules possesses antibacterial activity. Eur. J.
Biochem. 1996, 235, 516–525.
118. Vindrola, O.; Padrós, M.R.; Sterin-Prync, A.; Ase, A.; Finkielman, S.; Nahmod, V.
Proenkephalin system in human polymorphonuclear cells. Production and release of a novel
1.0-kD peptide derived from synenkephalin. J. Clin. Invest. 1990, 86, 531–537.
119. Metz-Boutigue, M.H.; Goumon, Y.; Strub, J.M.; Lugardon, K.; Aunis, D. Antimicrobial
chromogranins and proenkephalin-A-derived peptides. Ann. N. Y. Acad. Sci. 2003, 992, 168–178.
120. Shimizu, M.; Shigeri, Y.; Tatsu, Y.; Yoshikawa, S.; Yumoto, N. Enhancement of antimicrobial
activity of neuropeptide Y by N-terminal truncation. Antimicrob. Agents Chemother. 1998, 42,
2745–2746.
121. Hansen, C.J.; Burnell, K.K.; Brogden, K.A. Antimicrobial activity of Substance P
and Neuropeptide Y against laboratory strains of bacteria and oral microorganisms.
J. Neuroimmunol. 2006, 177, 215–218.
122. Allaker, R.P.; Zihni, C.; Kapas, S. An investigation into the antimicrobial effects of
adrenomedullin on members of the skin, oral, respiratory tract and gut microflora. FEMS
Immunol. Med. Microbiol. 1999, 23, 289–293.
123. Chu, H.; Pazgier, M.; Jung, G.; Nuccio, S.P.; Castillo, P.A.; de Jong, M.F.; Winter, M.G.;
Winter, S.E.; Wehkamp, J.; Shen, B.; et al. Human α-defensin 6 promotes mucosal innate
immunity through self-assembled peptide nanonets. Science 2012, 337, 477–481.
124. Wada, A.; Wong, P.F.; Hojo, H.; Hasegawa, M.; Ichinose, A.; Llanes, R.; Kubo, Y.; Senba, M.;
Ichinose, Y. Alarin but not its alternative-splicing form, GALP (Galanin-like peptide) has
antimicrobial activity. Biochem. Biophys. Res. Commun. 2013, 434, 223–227.
125. Brogden, K.A.; Guthmiller, J.M.; Salzet, M.; Zasloff, M. The nervous system and innate
immunity: The neuropeptide connection. Nat. Immunol. 2005, 6, 558–564.
126. Khemtémourian, L.; Killian, J.A.; Höppener, J.W.; Engel, M.F. Recent insights in islet amyloid
polypeptide-induced membrane disruption and its role in beta-cell death in type 2 diabetes
mellitus. Exp. Diabetes Res. 2008, 2008, 421287.
127. Shahnawaz, M.; Soto, C. Microcin amyloid fibrils A are reservoir of toxic oligomeric species.
J. Biol. Chem. 2012, 287, 11665–11676.
128. Domachowske, J.B.; Bonville, C.A.; Dyer, K.D.; Rosenberg, H.F. Evolution of antiviral
activity in the ribonuclease A gene superfamily: Evidence for a specific interaction between
eosinophil-derived neurotoxin (EDN/RNase 2) and respiratory syncytial virus. Nucleic Acids Res.
1998, 26, 5327–5332.
129. Pulido, D.; Torrent, M.; Andreu, D.; Nogués, M.V.; Boix, E. Two human host defense
ribonucleases against mycobacteria, the eosinophil cationic protein (RNase 3) and RNase 7.
Antimicrob. Agents Chemother. 2013, 57, 3797–805.
130. Huang, Y.C.; Lin, Y.M.; Chang, T.W.; Wu, S.J.; Lee, Y.S.; Chang, M.D.; Chen, C.; Wu, S.H.;
Liao, Y.D. The flexible and clustered lysine residues of human ribonuclease 7 are critical for
membrane permeability and antimicrobial activity. J. Biol. Chem. 2007, 282, 4626–4633.
131. Spencer, J.D.; Schwaderer, A.L.; Dirosario, J.D.; McHugh, K.M.; McGillivary, G.; Justice, S.S.;
Carpenter, A.R.; Baker, P.B.; Harder, J.; Hains, D.S. Ribonuclease 7 is a potent antimicrobial
peptide within the human urinary tract. Kidney Int. 2011, 80, 174–180.
132. Spencer, J.D.; Schwaderer, A.L.; Wang, H.; Bartz, J.; Kline, J.; Eichler, T.; DeSouza, K.R.;
Sims-Lucas, S.; Baker, P.; Hains, D.S. Ribonuclease 7, an antimicrobial peptide upregulated
during infection, contributes to microbial defense of the human urinary tract. Kidney Int. 2013,
83, 615–625.
133. Nielsen, K.L.; Dynesen, P.; Larsen, P.; Jakobsen, L.; Andersen, P.S.; Frimodt-Møller, N. Role of
urinary cathelicidin LL-37 and human β-defensin 1 in uncomplicated Escherichia coli urinary
tractinfections. Infect Immun. 2014, 82, 1572-1578.
134. Chromek, M.; Slamová, Z.; Bergman, P.; Kovács, L.; Podracká, L.; Ehrén, I.; Hökfelt, T.;
Gudmundsson, G.H.; Gallo, R.L.; Agerberth, B.; et al. The antimicrobial peptide cathelicidin
protects the urinary tract against invasive bacterial infection. Nat. Med. 2006, 12, 636–641.
135. Becknell, B.; Spencer, J.D.; Carpenter, A.R.; Chen, X.; Singh, A.; Ploeger, S.; Kline, J.;
Ellsworth, P.; Li, B.; Proksch, E.; et al. Expression and antimicrobial function of beta-defensin 1
in the lower urinary tract. PLoS One. 2013, 8, e77714.
136. Wiedłocha, A. Following angiogenin during angiogenesis: A journey from the cell surface to the
nucleolus. Arch. Immunol. Ther. Exp. (Warsz) 1999, 47, 299–305.
137. Avdeeva, S.V.; Chernukha, M.U.; Shaginyan, I.A.; Tarantul, V.Z.; Naroditsky, B.S. Human
angiogenin lacks specific antimicrobial activity. Curr. Microbiol. 2006, 53, 477–8.
138. Rudolph, B.; Podschun, R.; Sahly, H.; Schubert, S.; Schröder, J.M.; Harder, J. Identification of
RNase 8 as a novel human antimicrobial protein. Antimicrob. Agents Chemother. 2006, 50,
3194–3196.
139. Boix, E.; Torrent, M.; Sánchez, D.; Nogués, M.V. The antipathogen activities of eosinophil
cationic protein. Curr. Pharm. Biotechnol. 2008, 9, 141–152.
140. Hoffmann, H.J.; Olsen, E.; Etzerodt, M.; Madsen, P.; Thøgersen, H.C.; Kruse, T.; Celis, J.E.
Psoriasin binds calcium and is upregulated by calcium to levels that resemble those observed in
normal skin. J. Invest. Dermatol. 1994, 103, 370–375.
141. Porre, S.; Heinonen, S.; Mäntyjärvi, R.; Rytkönen-Nissinen, M.; Perola, O.; Rautiainen, J.;
Virtanen, T. Psoriasin, a calcium-binding protein with chemotactic properties is present in the
third trimester amniotic fluid. Mol. Hum. Reprod. 2005, 11, 87–92.
142. Ostergaard, M.; Rasmussen, H.H.; Nielsen, H.V.; Vorum, H.; Orntoft, T.F.; Wolf, H.; Celis, J.E.
Proteome profiling of bladder squamous cell carcinomas: Identification of markers that define
their degree of differentiation. Cancer Res. 1997, 57, 4111–4117.
143. Narushima, Y.; Unno, M.; Nakagawara, K.; Mori, M.; Miyashita, H.; Suzuki, Y.; Noguchi, N.;
Takasawa, S.; Kumagai, T.; Yonekura, H.; et al. Structure, chromosomal localization and
expression of mouse genes encoding type III Reg, RegIII α, RegIII β, RegIII γ. Gene 1997,
185, 159–168.
144. Feng, Y.; Huang, N.; Wu, Q.; Wang, B. HMGN2: A novel antimicrobial effector molecule of
human mononuclear leukocytes? J. Leukoc. Biol. 2005, 78, 1136–1141.
145. Domachowske, J.B.; Dyer, K.D.; Bonville, C.A.; Rosenberg, H.F. Recombinant human
eosinophil-derived neurotoxin/RNase 2 functions as an effective antiviral agent against
respiratory syncytial virus. J. Infect. Dis. 1998, 177, 1458–64.
146. Domachowske, J.B.; Dyer, K.D.; Adams, A.G.; Leto, T.L.; Rosenberg, H.F. Eosinophil cationic
protein/RNase 3 is another RNase A-family ribonuclease with direct antiviral activity. Nucleic
Acids Res. 1998, 26, 3358–63.
147. Shugars, D.C. Endogenous mucosal antiviral factors of the oral cavity. J. Infect. Dis. 1999, 179,
S431–S435.
148. Wang, W.; Owen, S.M.; Rudolph, D.L.; Cole, A.M.; Hong, T.; Waring, A.J.; Lal, R.B.;
Lehrer, R.I. Activity of alpha- and theta-defensins against primary isolates of HIV-1. J. Immunol.
2004, 173, 515–520.
149. Dugan, A.S.; Maginnis, M.S.; Jordan, J.A.; Gasparovic, M.L.; Manley, K.; Page, R.; Williams, G.;
Porter, E.; O'Hara, B.A.; Atwood, W.J. Human alpha-defensins inhibit BK virus infection by
aggregating virions and blocking binding to host cells. J. Biol. Chem. 2008, 283, 31125–31132.
150. Quiñones-Mateu, M.E.; Lederman, M.M.; Feng, Z.; Chakraborty, B.; Weber, J.; Rangel, H.R.;
Marotta, M.L.; Mirza, M.; Jiang, B.; Kiser, P.; et al. Human epithelial beta-defensins 2 and 3
inhibit HIV-1 replication. AIDS 2003, 17, F39–F48.
151. Bergman, P.; Walter-Jallow, L.; Broliden, K.; Agerberth, B.; Söderlund, J. The antimicrobial
peptide LL-37 inhibits HIV-1 replication. Curr. HIV Res. 2007, 5, 410–415.
152. Wang, G.; Watson, K.M.; Buckheit RW, Jr. Anti-human immunodeficiency virus type 1
activities of antimicrobial peptides derived from human and bovine cathelicidins. Antimicrob.
153. Groot, F.; Sanders, R.W.; ter Brake, O.; Nazmi, K.; Veerman, E.C.; Bolscher, J.G.; Berkhout, B.
Histatin 5-derived peptide with improved fungicidal properties enhances human immunodeficiency
virus type 1 replication by promoting viral entry. J. Virol. 2006, 80, 9236–9243.
154. Wang, G. Natural antimicrobial peptides as promising anti-HIV candidates. Curr. Topics Peptide
Protein Res. 2012, 13, 93–110.
155. López-Garcí a, B.; Lee, P.H.; Yamasaki, K.; Gallo, R.L. Anti-fungal activity of cathelicidins and
their potential role in Candida albicans skin infection. J. Invest. Dermatol. 2005, 125, 108–115.
156. Den Hertog, A.L.; van Marle, J.; Veerman, E.C.; Valentijn-Benz, M.; Nazmi, K.; Kalay, H.;
Grün, C.H.; Van’t Hof, W.; Bolscher, J.G.; Nieuw Amerongen, A.V. The human cathelicidin
peptide LL-37 and truncated variants induce segregation of lipids and proteins in the plasma
membrane of Candida albicans. Biol. Chem. 2006, 387, 1495–1502.
157. Dabirian, S.; Taslimi, Y.; Zahedifard, F.; Gholami, E.; Doustdari, F.; Motamedirad, M.;
Khatami, S.; Azadmanesh, K.; Nylen, S.; Rafati, S. Human neutrophil peptide-1 (HNP-1): A new
anti-leishmanial drug candidate. PLoS Negl. Trop. Dis. 2013, 7, e2491.
158. Söbirk, S.K.; Mörgelin, M.; Egesten, A.; Bates, P.; Shannon, O.; Collin, M. Human chemokines
as antimicrobial peptides with direct parasiticidal effect on Leishmania mexicana in vitro. PLoS
One 2013, 8, e58129.
159. Rico-Mata, R.; De Leon-Rodriguez, L.M.; Avila, E.E. Effect of antimicrobial peptides derived
from human cathelicidin LL-37 on Entamoeba histolytica trophozoites. Exp. Parasitol. 2013,
133, 300–306.
160. Love, M.S.; Millholland, M.G.; Mishra, S.; Kulkarni, S.; Freeman, K.B.; Pan, W.; Kavash, R.W.;
Costanzo, M.J.; Jo, H.; Daly, T.M.; et al. Platelet factor 4 activity against P. falciparum and its
translation to nonpeptidic mimics as antimalarials. Cell Host Microbe. 2012, 12, 815–823.
161. Baker, M.A.; Maloy, W.L.; Zasloff, M.; Jacob, L.S. Anticancer efficacy of Magainin2 and
analogue peptides. Cancer Res. 1993, 53, 3052–3057.
162. Winder, D.; Günzburg, W.H.; Erfle, V.; Salmons, B. Expression of antimicrobial peptides has an
antitumour effect in human cells. Biochem. Biophys. Res. Commun. 1998, 242, 608–612.
163. Lichtenstein, A.; Ganz, T.; Selsted, M.E.; Lehrer, R.I. In vitro tumor cell cytolysis mediated by
peptide defensins of human and rabbit granulocytes. Blood 1986, 68, 1407–1410.
164. Gaspar, D.; Veiga, A.S.; Castanho, M.A. From antimicrobial to anticancer peptides. A review.
Front. Microbiol. 2013, 4, 294.
165. Riedl, S.; Zweytick, D.; Lohner, K. Membrane-active host defense peptides—Challenges and
perspectives for the development of novel anticancer drugs. Chem. Phys. Lipids 2011, 164,
766–781.
166. Hoskin, D.W.; Ramamoorthy, A. Studies on anticancer activities of antimicrobial peptides.
Biochim. Biophys. Acta 2008, 1778, 357–375.
167. Kishi, A.; Takamori, Y.; Ogawa, K.; Takano, S.; Tomita, S.; Tanigawa, M.; Niman, M.; Kishida, T.;
Fujita, S. Differential expression of granulysin and perforin by NK cells in cancer patients and
correlation of impaired granulysin expression with progression of cancer. Cancer Immunol.
Immunother. 2002, 50, 604–614.
168. Sekiguchi, N.; Asano, N.; Ito, T.; Momose, K.; Momose, M.; Ishida, F. Elevated serum
granulysin and its clinical relevance in mature NK-cell neoplasms. Int. J. Hematol. 2012, 96,
461–468.
169. Xu, N.; Wang, Y.S.; Pan, W.B.; Xiao, B.; Wen, Y.J.; Chen, X.C.; Chen, L.J.; Deng, H.X.;
You, J.; Kan, B.; et al. Human alpha-defensin-1 inhibits growth of human lung adenocarcinoma
xenograft in nude mice. Mol. Cancer Ther. 2008, 7, 1588–1597.
170. Mizukawa, N.; Sugiyama, K.; Fukunaga, J.; Ueno, T.; Mishima, K.; Takagi, S.; Sugahara, T.
Defensin-1, a peptide detected in the saliva of oral squamous cell carcinoma patients. Anticancer
Res. 1998, 18, 4645–4649.
171. Müller, C.A.; Markovic-Lipkovski, J.; Klatt, T.; Gamper, J.; Schwarz, G.; Beck, H.; Deeg, M.;
Kalbacher, H.; Widmann, S.; Wessels, J.T.; et al. Human alpha-defensins HNPs-1, -2, and -3 in
renal cell carcinoma: Influences on tumor cell proliferation. Am. J. Pathol. 2002, 160, 1311–1324.
172. Sun, C.Q.; Arnold, R.; Fernandez-Golarz, C.; Parrish, A.B.; Almekinder, T.; He, J.; Ho, S.M.;
Svoboda, P.; Pohl, J.; Marshall, F.F.; et al. Human beta-defensin-1, a potential chromosome 8p
tumor suppressor: Control of transcription and induction of apoptosis in renal cell carcinoma.
Cancer Res. 2006, 66, 8542–8549.
173. Bullard, R.S.; Gibson, W.; Bose, S.K.; Belgrave, J.K.; Eaddy, A.C.; Wright, C.J.;
Hazen-Martin, D.J.; Lage, J.M.; Keane, T.E.; Ganz, T.A.; et al. Functional analysis of the host
defense peptide Human Beta Defensin-1: New insight into its potential role in cancer. Mol.
Immunol. 2008, 45, 839–848.
174. Winter, J.; Pantelis, A.; Reich, R.; Martini, M.; Kraus, D.; Jepsen, S.; Allam, J.P.; Novak, N.;
Wenghoefer, M. Human beta-defensin-1, -2, and -3 exhibit opposite effects on oral squamous
cell carcinoma cell proliferation. Cancer Invest. 2011, 29, 196–201.
175. Wu, W.K.; Wang, G.; Coffelt, S.B.; Betancourt, A.M.; Lee, C.W.; Fan, D.; Wu, K.; Yu, J.;
Sung, J.J.; Cho, C.H. Emerging roles of the host defense peptide LL-37 in human cancer and its
potential therapeutic applications. Int. J. Cancer. 2010, 127, 1741–1747.
176. van den Broek, I.; Sparidans, R.W.; Engwegen, J.Y.; Cats, A.; Depla, A.C.; Schellens, J.H.;
Beijnen, J.H. Evaluation of human neutrophil peptide-1, -2 and -3 as serum markers for
colorectal cancer. Cancer Biomark. 2010, 7, 109–115.
177. Albrethsen, J.; Bøgebo, R.; Gammeltoft, S.; Olsen, J.; Winther, B.; Raskov, H. Upregulated
expression of human neutrophil peptides 1, 2 and 3 (HNP 1–3) in colon cancer serum and
tumours: A biomarker study. BMC Cancer 2005, 5, 8.
178. Albrethsen, J.; Møller, C.H.; Olsen, J.; Raskov, H.; Gammeltoft, S. Human neutrophil peptides 1,
2 and 3 are biochemical markers for metastatic colorectal cancer. Eur. J. Cancer 2006, 42,
3057–3064.
179. Li, X.; Li, Y.; Han, H.; Miller, D.W.; Wang, G. Solution structures of human LL-37 fragments
and NMR-based identification of a minimal membrane-targeting antimicrobial and anticancer
region. J. Am. Chem. Soc. 2006, 128, 5776–5785.
180. Ren, S.X.; Shen, J.; Cheng, A.S.; Lu, L.; Chan, R.L.; Li, Z.J.; Wang, X.J.; Wong, C.C.;
Zhang, L.; Ng, S.S.; et al. FK-16 derived from the anticancer peptide LL-37 induces caspase-
independent apoptosis and autophagic cell death in colon cancer cells. PLoS One 2013, 8,
e63641.
181. Zhou, J.; Shi, J.; Hou, J.; Cao, F.; Zhang, Y.; Rasmussen, J.T.; Heegaard, C.W.; Gilbert, G.E.
Phosphatidylserine exposure and procoagulant activity in acute promyelocytic leukemia.
J. Thromb. Haemost. 2010, 8, 773–782.
182. Wang, G. Structures of human host defense cathelicidin LL-37 and its smallest antimicrobial
peptide KR-12 in lipid micelles. J. Biol. Chem. 2008, 283, 32637–32643.
183. Svensson, D.; Westman, J.; Wickström, C.; Jönsson, D.; Herwald, H.; Nilsson, B.O. Human
endogenous peptide p33 inhibits detrimental effects of LL-37 on osteoblast viability.
J. Periodontal Res. 2014, doi:10.1111/jre.12184.
184. Hiemstra, T.F.; Charles, P.D.; Gracia, T.; Hester, S.S.; Gatto, L.; Al-Lamki, R.; Floto, R.A.;
Su, Y.; Skepper, J.N.; Lilley, K.S.; et al. Human Urinary Exosomes as Innate Immune Effectors.
J. Am. Soc. Nephrol. 2014, in press.
185. Yung, S.C.; Murphy, P.M. Antimicrobial chemokines. Front Immunol. 2012; 3, 276.
186. Yang, D.; Chen, Q.; Schmidt, A.P.; Anderson, G.M.; Wang, J.M.; Wooters, J.; Oppenheim, J.J.;
Chertov, O. LL-37, the neutrophil granule- and epithelial cell-derived cathelicidin, utilizes
formyl peptide receptor-like 1 (FPRL1) as a receptor to chemoattract human peripheral blood
neutrophils, monocytes, and T cells. J. Exp. Med. 2000, 192, 1069–1074.
187. Ciornei, C.D.; Tapper, H.; Bjartell, A.; Sternby, N.H.; Bodelsson, M. Human antimicrobial
peptide LL-37 is present in atherosclerotic plaques and induces death of vascular smooth muscle
cells: A laboratory study. BMC Cardiovasc. Disord. 2006, 6, 49.
188. Barlow, P.G.; Li, Y.; Wilkinson, T.S.; Bowdish, D.M.; Lau, Y.E.; Cosseau, C.; Haslett, C.;
Simpson, A.J.; Hancock, R.E.; Davidson, D.J. The human cationic host defense peptide LL-37
mediates contrasting effects on apoptotic pathways in different primary cells of the innate
immune system. J. Leukoc. Biol. 2006, 80, 509–520.
189. Aarbiou, J.; Tjabringa, G.S.; Verhoosel, R.M.; Ninaber, D.K.; White, S.R.; Peltenburg, L.T.;
Rabe, K.F.; Hiemstra, P.S. Mechanisms of cell death induced by the neutrophil antimicrobial
peptides alpha-defensins and LL-37. Inflamm. Res. 2006, 55, 119–127.
190. Okumura, K.; Itoh, A.; Isogai, E.; Hirose, K.; Hosokawa, Y.; Abiko, Y.; Shibata, T.; Hirata, M.;
Isogai, H. C-terminal domain of human CAP18 antimicrobial peptide induces apoptosis in oral
squamous cell carcinoma SAS-H1 cells. Cancer Lett. 2004, 212, 185–194.
191. Ciornei, C.D.; Egesten, A.; Bodelsson, M. Effects of human cathelicidin antimicrobial peptide
LL-37 on lipopolysaccharide-induced nitric oxide release from rat aorta in vitro. Acta
Anaesthesiol. Scand. 2003, 47, 213–220.
192. Chamorro, C.I.; Weber, G.; Grönberg, A.; Pivarcsi, A.; Ståhle, M. The human antimicrobial
peptide LL-37 suppresses apoptosis in keratinocytes. J. Invest. Dermatol. 2009, 129, 937–944.
193. Nagaoka, I.; Tamura, H.; Hirata, M. An antimicrobial cathelicidin peptide, human CAP18/
LL-37, suppresses neutrophil apoptosis via the activation of formyl-peptide receptor-like 1 and
P2X7. J. Immunol. 2006, 176, 3044–3052.
194. Lee, W.Y.; Savage, J.R.; Zhang, J.; Jia, W.; Oottamasathien, S.; Prestwich, G.D. Prevention of
anti-microbial peptide LL-37-induced apoptosis and ATP release in the urinary bladder by a
modified glycosaminoglycan. PLoS One 2013, 8, e77854.
195. Suzuki, K.; Murakami, T.; Kuwahara-Arai, K.; Tamura, H.; Hiramatsu, K.; Nagaoka, I. Human
anti-microbial cathelicidin peptide LL-37 suppresses the LPS-induced apoptosis of endothelial
cells. Int. Immunol. 2011, 23, 185–193.
196. Ren, S.X.; Cheng, A.S.; To, K.F.; Tong, J.H.; Li, M.S.; Shen, J.; Wong, C.C.; Zhang, L.;
Chan, R.L.; Wang, X.J.; et al. Host immune defense peptide LL-37 activates
caspase-independent apoptosis and suppresses colon cancer. Cancer Res. 2012, 72, 6512–6523.
197. Oudhoff, M.J.; Blaauboer, M.E.; Nazmi, K.; Scheres, N.; Bolscher, J.G.; Veerman, E.C. The role
of salivary histatin and the human cathelicidin LL-37 in wound healing and innate immunity.
Biol. Chem. 2010, 391, 541–548.
198. Wang, G. NMR of membrane-associated peptides and proteins. Curr. Protein Pept. Sci. 2008, 9,
50–69.
199. Wang, G. NMR of membrane proteins. In ―Advances in Protein and Peptide Sciences‖ (edited by
Dunn BM). Bentham Sci. 2013, 1, 128–188.
200. Nguyen, L.T.; Haney, E.F.; Vogel, H.J. The expanding scope of antimicrobial peptide structures
and their modes of action. Trends Biotechnol. 2011, 29, 464–472.
201. Wang, G.; Li, X.; Zasloff, M. A Database View of Natural Antimicrobial Peptides:
Nomenclature, Classification and Amino acid Sequence Analysis. In Antimicrobial Peptides:
Discovery, Design and Novel Therapeutic Strategies; Wang, G., Ed.; CABI: Wallingford, UK,
2010; pp. 1–21.
202. Rose, P.W.; Bi, C.; Bluhm, W.F.; Christie, C.H.; Dimitropoulos, D.; Dutta, S.; Green, R.K.;
Goodsell, D.S.; Prlic, A.; Quesada, M.; et al. The RCSB Protein Data Bank: New resources for
research and education. Nucleic Acids Res. 2013, 41, D475–D482.
203. Xu, T.; Levitz, S.M.; Diamond, R.D.; Oppenheim, F.G. Anticandidal activity of major human
salivary histatins. Infect. Immun. 1991, 59, 2549–2554.
204. Raj, P.A.; Edgerton, M.; Levine, M.J. Salivary histatin 5: Dependence of sequence, chain length,
and helical conformation for candidacidal activity. J. Biol. Chem. 1990, 265, 3898–3905.
205. Rothstein, D.M.; Spacciapoli, P.; Tran, L.T.; Xu, T.; Roberts, F.D.; Dalla Serra, M.;
Buxton, D.K.; Oppenheim, F.G.; Friden, P. Anticandida activity is retained in P-113, a
12-amino-acid fragment of histatin 5. Antimicrob. Agents Chemother. 2001, 45, 1367–1373.
206. Zuo, Y.; Xu, T.; Troxler, R.F.; Li, J.; Driscoll, J.; Oppenheim, F.G. Recombinant histatins:
Functional domain duplication enhances candidacidal activity. Gene 1995, 161, 87–91.
207. Situ, H.; Tsai, H.; Bobek, L.A. Construction and characterization of human salivary histatin-5
multimers. J. Dent. Res. 1999, 78, 690–698.
208. Situ, H.; Balasubramanian, S.V.; Bobek, L.A. Role of alpha-helical conformation of histatin-5 in
candidacidal activity examined by proline variants. Biochim. Biophys. Acta 2000, 1475,
377–382.
209. Melino, S.; Rufini, S.; Sette, M.; Morero, R.; Grottesi, A.; Paci, M.; Petruzzelli, R. Zn2+ ions
selectively induce antimicrobial salivary peptide histatin-5 to fuse negatively charged vesicles.
Identification and characterization of a zinc-binding motif present in the functional domain.
Biochemistry 1999, 38, 9626–9633.
210. Rydengård, V.; Andersson Nordahl, E.; Schmidtchen, A. Zinc potentiates the antibacterial effects
of histidine-rich peptides against Enterococcus faecalis. FEBS J. 2006, 273, 2399–2406.
211. Melino, S.; Gallo, M.; Trotta, E.; Mondello, F.; Paci, M.; Petruzzelli, R. Metal-binding and
nuclease activity of an antimicrobial peptide analogue of the salivary histatin 5. Biochemistry
2006, 45, 15373–15383.
212. Wang, G. Structural studies of antimicrobial peptides provide insight into their mechanisms of
action. In Antimicrobial Peptides: Discovery, Design and Novel Therapeutic Strategies;
Wang, G., Ed.; CABI: Wallingford, UK, 2010; pp. 141–168.
213. Xhindoli, D.; Pacor, S.; Guida, F.; Antcheva, N.; Tossi, A. Native oligomerization determines the
mode of action and biological activities of human cathelicidin LL-37. Biochem. J. 2014, 457,
263–275.
214. Wang, G.; Elliott, M.; Cogen, A.L.; Ezell, E.L.; Gallo, R.L.; Hancock, R.E.W. Structure,
dynamics, antimicrobial and immune modulatory activities of human LL-23 and its single
residue variants mutated based on homologous primate cathelicidins. Biochemistry 2012, 51,
653–664.
215. Paulmann, M.; Arnold, T.; Linke, D.; Özdirekcan, S.; Kopp, A.; Gutsmann, T.; Kalbacher, H.;
Wanke, I.; Schuenemann, V.J.; Habeck, M.; et al. Structure-activity analysis of the
dermcidin-derived peptide DCD-1L, an anionic antimicrobial peptide present in human sweat.
J. Biol. Chem. 2012, 287, 8434–8443]
216. Song, C.; Weichbrodt, C.; Salnikov, E.S.; Dynowski, M.; Forsberg, B.O.; Bechinger, B.;
Steinem, C.; de Groot, B.L.; Zachariae, U.; Zeth, K. Crystal structure and functional mechanism
of a human antimicrobial membrane channel. Proc. Natl. Acad. Sci. USA 2013, 110, 4586–4591.
217. Jung, H.H.; Yang, S.T.; Sim, J.Y.; Lee, S.; Lee, J.Y.; Kim, H.H.; Shin, S.Y.; Kim, J.I. Analysis
of the solution structure of the human antibiotic peptide dermcidin and its interaction with
phospholipid vesicles. BMB Rep. 2010, 43, 362–368.
218. Koradi, R.; Billeter, M.; Wüthrich, K. MOLMOL: A program for display and analysis of
macromolecular structures. J. Mol. Graph. 1996, 14, 51–55.
219. Anderson, D.H.; Sawaya, M.R.; Cascio, D.; Ernst, W.; Modlin, R.; Krensky, A.; Eisenberg, D.
Granulysin crystal structure and a structure-derived lytic mechanism. J. Mol. Biol. 2003, 325,
355–365.
220. Peña, S.V.; Krensky, A.M. Granulysin, a new human cytolytic granule-associated protein with
possible involvement in cell-mediated cytotoxicity. Semin. Immunol. 1997, 9, 117–125.
221. Bruhn, H. A short guided tour through functional and structural features of saposin-like proteins.
Biochem. J. 2005, 389 (Pt 2), 249–257.
222. da Silva, A.P.; Unks, D.; Lyu, S.C.; Ma, J.; Zbozien-Pacamaj, R.; Chen, X.; Krensky, A.M.;
Clayberger, C. In vitro and in vivo antimicrobial activity of granulysin-derived peptides against
Vibrio cholerae. J Antimicrob Chemother. 2008, 61, 1103–1109.
223. McInturff, J.E.; Wang, S.J.; Machleidt, T.; Lin, T.R.; Oren, A.; Hertz, C.J.; Krutzik, S.R.;
Hart, S.; Zeh, K.; Anderson, D.H.; et al. Granulysin-derived peptides demonstrate antimicrobial
and anti-inflammatory effects against Propionibacterium. acnes. J. Invest. Dermatol. 2005, 125,
256–263.
224. Ericksen, B.; Wu, Z.; Lu, W.; Lehrer, R.I. Antibacterial activity and specificity of the six human
α-defensins. Antimicrob. Agents Chemother. 2005, 49, 269–275.
225. Schibli, D.J.; Hunter, H.N.; Aseyev, V.; Starner, T.D.; Wiencek, J.M.; McCray PB, Jr.;
Tack, B.F.; Vogel, H.J. The solution structures of the human beta-defensins lead to a better
understanding of the potent bactericidal activity of HBD3 against Staphylococcus aureus. J. Biol.
Chem. 2002, 277, 8279–8289.
226. Schroeder, B.O.; Wu, Z.; Nuding, S.; Groscurth, S.; Marcinowski, M.; Beisner, J.; Buchner, J.;
Schaller, M.; Stange, E.F.; Wehkamp, J. Reduction of disulphide bonds unmasks potent
antimicrobial activity of human β-defensin 1. Nature 2011, 469, 419–23.
227. Keizer, D.W.; Crump, M.P.; Lee, T.W.; Slupsky, C.M.; Clark-Lewis, I.; Sykes, B.D. Human CC
chemokine I-309, structural consequences of the additional disulfide bond. Biochemistry 2000,
39, 6053–6059.
228. Blaszczyk, J.; Coillie, E.V.; Proost, P.; Damme, J.V.; Opdenakker, G.; Bujacz, G.D.;
Wang, J.M.; Ji, X. Complete crystal structure of monocyte chemotactic protein-2, a CC
chemokine that interacts with multiple receptors. Biochemistry 2000, 39, 14075–14081.
229. Crump, M.P.; Rajarathnam, K.; Kim, K.S.; Clark-Lewis, I.; Sykes, B.D. Solution structure of
eotaxin, a chemokine that selectively recruits eosinophils in allergic inflammation. J. Biol. Chem.
1998, 273, 22471–22479.
230. Love, M.; Sandberg, J.L.; Ziarek, J.J.; Gerarden, K.P.; Rode, R.R.; Jensen, D.R.;
McCaslin, D.R.; Peterson, F.C.; Veldkamp, C.T. Solution structure of CCL21 and identification
of a putative CCR7 binding site. Biochemistry 2012, 51, 733–735.
231. Jansma, A.L.; Kirkpatrick, J.P.; Hsu, A.R.; Handel, T.M.; Nietlispach, D. NMR analysis of the
structure, dynamics, and unique oligomerization properties of the chemokine CCL27. J. Biol.
Chem. 2010, 285, 14424–37
232. Veldkamp, C.T.; Ziarek, J.J.; Su, J.; Basnet, H.; Lennertz, R.; Weiner, J.J.; Peterson, F.C.;
Baker, J.E.; Volkman, B.F. Monomeric structure of the cardioprotective chemokine
SDF-1/CXCL12. Protein Sci. 2009, 18, 1359–1369.
233. Hoover, D.M.; Boulegue, C.; Yang, D.; Oppenheim, J.J.; Tucker, K.; Lu, W.; Lubkowski, J. The
structure of human macrophage inflammatory protein-3alpha/CCL20. Linking antimicrobial and
CC chemokine receptor-6-binding activities with human beta-defensins. J. Biol. Chem. 2002,
277, 37647–37654.
234. Barinka, C.; Prahl, A.; Lubkowski, J. Structure of human monocyte chemoattractant protein 4
(MCP-4/CCL13). Acta. Crystallogr. D Biol. Crystallogr. 2008, 64(Pt 3), 273–278
235. Kim, K.S.; Clark-Lewis, I.; Sykes, B.D. Solution structure of GRO/melanoma growth stimulatory
activity determined by 1H-NMR spectroscopy. J. Biol. Chem. 1994, 269, 32909–32915.
236. Swaminathan, G.J.; Holloway, D.E.; Colvin, R.A.; Campanella, G.K.; Papageorgiou, A.C.;
Luster, A.D.; Acharya, K.R. Crystal structures of oligomeric forms of the IP-10/CXCL-10
chemokine. Structure 2003, 11, 521–532.
237. Yung, S.C.; Parenti, D.; Murphy, P.M. Host chemokines bind to Staphylococcus aureus and
stimulate protein A release. J. Biol. Chem. 2011, 286, 5069–5077.
238. Nguyen, L.T.; Kwakman, P.H.; Chan, D.I.; Liu, Z.; de Boer, L.; Zaat, S.A.; Vogel, H.J.
Exploring platelet chemokine antimicrobial activity: Nuclear magnetic resonance backbone
dynamics of NAP-2 and TC-1. Antimicrob. Agents Chemother. 2011, 55, 2074–2083.
239. Wang, G.; Epand, R.F.; Mishra, B.; Lushnikova, T.; Thomas, V.C.; Bayles, K.W.; Epand, R.M.
Decoding the functional roles of cationic side chains of the major antimicrobial region of human
cathelicidin LL-37. Antimicrob. Agents Chemother. 2012, 56, 845–856.
240. Lequin, O.; Thüring, H.; Robin, M.; Lallemand, J.Y. Three-dimensional solution structure of
human angiogenin determined by 1H,15N-NMR spectroscopy--characterization of histidine
protonation states and pKa values. Eur. J. Biochem. 1997, 250, 712–726.
241. Wang, H.; Schwaderer, A.L.; Kline, J.; Spencer, J.D.; Kline, D.; Hains, D.S. Contribution of
structural domains to the activity of ribonuclease 7 against uropathogenic bacteria. Antimicrob.
242. Torrent, M.; Pulido, D.; Valle, J.; Nogués, M.V.; Andreu, D.; Boix, E. Ribonucleases as a
host-defence family: Evidence of evolutionarily conserved antimicrobial activity at the
N-terminus. Biochem. J. 2013, 456, 99–108.
243. Boix, E.; Salazar, V.A.; Torrent, M.; Pulido, D.; Nogués, M.V.; Moussaoui, M. Structural
determinants of the eosinophil cationic protein antimicrobial activity. Biol. Chem. 2012, 393,
801–815.
244. Spencer, J.D.; Schwaderer, A.L.; Eichler, T.; Wang, H.; Kline, J.; Justice, S.S.; Cohen, D.M.;
Hains, D.S. An endogenous ribonuclease inhibitor regulates the antimicrobial activity of
ribonuclease 7 in the human urinary tract. Kidney Int. 2013, 85, 1179–1191.
245. Lehotzky, R.E.; Partch, C.L.; Mukherjee, S.; Cash, H.L.; Goldman, W.E.; Gardner, K.H.;
Hooper, L.V. Molecular basis for peptidoglycan recognition by a bactericidal lectin. Proc. Natl.
Acad. Sci. USA 2010, 107, 7722–7727.
246. Mukherjee, S.; Zheng, H.; Derebe, M.G.; Callenberg, K.M.; Partch, C.L.; Rollins, D.;
Propheter, D.C.; Rizo, J.; Grabe, M.; Jiang, Q.X.; et al. Antibacterial membrane attack by a pore-
forming intestinal C-type lectin. Nature 2013, 505, 103–107.
247. De Leeuw, E.; Li, C.; Zeng, P.; Li, C.; Diepeveen-de Buin, M.; Lu, W.Y.; Breukink, E.; Lu, W.
Functional interaction of human neutrophil peptide-1 with the cell wall precursor lipid II. FEBS
Lett. 2010, 584, 1543–1548.
248. Böhling, A.; Hagge, S.O.; Roes, S.; Podschun, R.; Sahly, H.; Harder, J.; Schröder, J.M.;
Grötzinger, J.; Seydel, U.; Gutsmann, T. Lipid-specific membrane activity of human
beta-defensin-3. Biochemistry 2006, 45, 5663–5670.
249. Sass, V.; Schneider, T.; Wilmes, M.; Körner, C.; Tossi, A.; Novikova, N.; Shamova, O.;
Sahl, H.G. Human beta-defensin 3 inhibits cell wall biosynthesis in Staphylococci. Infect.
Immun. 2010, 78, 2793–2800.
250. Schmitt, P.; Wilmes, M.; Pugnière, M.; Aumelas, A.; Bachère, E.; Sahl, H.G.; Schneider, T.;
Destoumieux-Garzón, D. Insight into invertebrate defensin mechanism of action: Oyster
defensins inhibit peptidoglycan biosynthesis by binding to lipid II. Biol. Chem. 2010, 285,
29208–29216.
251. Bierbaum, G.; Sahl, H.G. Lantibiotics: Mode of action, biosynthesis and bioengineering. Curr.
Pharm. Biotechnol. 2009, 10, 2–18.
252. Schneider, T.; Kruse, T.; Wimmer, R.; Wiedemann, I.; Sass, V.; Pag, U.; Jansen, A.;
Nielsen, A.K.; Mygind, P.H.; Raventós, D.S.; et al. Plectasin, a fungal defensin, targets the
bacterial cell wall precursor Lipid II. Science 2010, 328, 1168–1172.
253. Derouaux, A.; Turk, S.; Olrichs, N.K.; Gobec, S.; Breukink, E.; Amoroso, A.; Offant, J.;
Bostock, J.; Mariner, K.; Chopra, I.; et al. Small molecule inhibitors of peptidoglycan synthesis
targeting the lipid II precursor. Biochem. Pharmacol. 2011, 81, 1098–1105.
254. Varney, K.M.; Bonvin, A.M.; Pazgier, M.; Malin, J.; Yu, W.; Ateh, E.; Oashi, T.; Lu, W.;
Huang, J.; Diepeveen-de Buin, M.; et al. Turning Defense into Offense: Defensin Mimetics as
Novel Antibiotics Targeting Lipid II. PLoS Pathog. 2013, 9, e1003732.
255. Poon, I.K.h.; Baxter, A.A.; Lay, F.T.; Mills, G.D.; Adda, C.G.; Payne, J.A.; Phan, T.K.;
Ryan, G.F.; White, J.A.; Veneer, P.K.; et al. Phosphoinositide-mediated oligomerization of a
defensin induces cell lysis. Elife 2014, 3, e01808.
256. Silva, P.M.; Gonçalves, S.; Santos, N.C. Defensins: Antifungal lessons from eukaryotes. Front.
Microbiol. 2014, 5, 97.
257. Sagaram, U.S.; El-Mounadi, K.; Buchko, G.W.; Berg, H.R.; Kaur, J.; Pandurangi, R.S.; Smith,
T.J.; Shah, D.M. Structural and functional studies of a phosphatidic acid-binding antifungal plant
defensin MtDef4: identification of an RGFRRR motif governing fungal cell entry. PLoS One.
2013, 8, e82485.
258. Miki, T.; Holst, O.; Hardt, W.D. The bactericidal activity of the C-type lectin RegIIIβ against
Gram-negative bacteria involves binding to lipid A. J. Biol. Chem. 2012, 287, 34844–34855.
259. Formanek, H. A three dimensional model of the digestion of peptidoglycan by lysozyme.
Biophys. Struct. Mech. 1977, 4, 1–14.
260. Ma, G.; Greenwell-Wild, T.; Lei, K.; Jin, W.; Swisher, J.; Hardegen, N.; Wild, C.T.; Wahl, S.M.
Secretory leukocyte protease inhibitor binds to annexin II, a cofactor for macrophage HIV-1
infection. J. Exp. Med. 2004, 200, 1337–1346.
261. Oren, Z.; Lerman, J.C.; Gudmundsson, G.H.; Agerberth, B.; Shai, Y. Structure and organization
of the human antimicrobial peptide LL-37 in phospholipid membranes: Relevance to the
molecular basis for its non-cell-selective activity. Biochem. J. 1999, 341 (Pt. 3), 501–513.
262. Lee, C.C.; Sun, Y.; Qian, S.; Huang, H.W. Transmembrane pores formed by human
antimicrobial peptide LL-37. Biophys. J. 2011, 100, 1688–1696.
263. Maisetta, G.; Vitali, A.; Scorciapino, M.A.; Rinaldi, A.C.; Petruzzelli, R.; Brancatisano, F.L.;
Esin, S.; Stringaro, A.; Colone, M.; Luzi, C.; et al. pH-dependent disruption of Escherichia coli
ATCC 25922 and model membranes by the human antimicrobial peptides hepcidin 20 and 25.
FEBS J. 2013, 280, 2842–2854.
264. Natividad, J.M.; Hayes, C.L.; Motta, J.P.; Jury, J.; Galipeau, H.J.; Philip, V.;
Garcia-Rodenas, C.L.; Kiyama, H.; Bercik, P.; Verdu, E.F. Differential Induction of
Antimicrobial REGIII by the Intestinal Microbiota and Bifidobacterium breve NCC2950. Appl.
Environ. Microbiol. 2013, 79, 7745–7754.
265. Parker, M.W.; Feil, S.C. Pore-forming protein toxins: From structure to function. Prog. Biophys.
Mol. Biol. 2005, 88, 91–142.
266. Birck, C.; Damian, L.; Marty-Detraves, C.; Lougarre, A.; Schulze-Briese, C.; Koehl, P.;
Fournier, D.; Paquereau, L.; Samama, J.P. A new lectin family with structure similarity to
actinoporins revealed by the crystal structure of Xerocomus chrysenteron lectin XCL. J. Mol.
Biol. 2004, 344, 1409–1420.
267. Mechaly, A.E.; Bellomio, A.; Gil-Cartón, D.; Morante, K.; Valle, M.; González-Mañas, J.M.;
Guérin, D.M. Structural insights into the oligomerization and architecture of eukaryotic
membrane pore-forming toxins. Structure 2011, 19, 181–191.
268. Miller, K.W.; Evans, R.J.; Eisenberg, S.P.; Thompson, R.C. Secretory leukocyte protease
inhibitor binding to mRNA and DNA as a possible cause of toxicity to Escherichia coli.
J. Bacteriol. 1989, 171, 2166–2172.
269. den Hertog, A.L.; van Marle, J.; van Veen, H.A.; Van't Hof, W.; Bolscher, J.G.; Veerman, E.C.;
Nieuw Amerongen, A.V. Candidacidal effects of two antimicrobial peptides: Histatin 5 causes
small membrane defects, but LL-37 causes massive disruption of the cell membrane. Biochem. J.
2005, 388, 689–695
270. Gyurko, C.; Lendenmann, U.; Troxler, R.F.; Oppenheim, F.G. Candida albicans mutants
deficient in respiration are resistant to the small cationic salivary antimicrobial peptide histatin 5.
Antimicrob. Agents Chemother. 2000, 44, 348–354.
271. Gyurko, C.; Lendenmann, U.; Helmerhorst, E.J.; Troxler, R.F.; Oppenheim, F.G. Killing of
Candida albicans by histatin 5: Cellular uptake and energy requirement. Antonie. Van
Leeuwenhoek. 2001, 79, 297–309.
272. Li, X.S.; Sun, J.N.; Okamoto-Shibayama, K.; Edgerton, M. Candida albicans cell wall ssa
proteins bind and facilitate import of salivary histatin 5 required for toxicity. J. Biol. Chem. 2006,
281, 22453–22463
273. Helmerhorst, E.J.; Breeuwer, P.; Van’t Hof, W.; Walgreen-Weterings, E.; Amerongen, A.V.;
Abee, T. The cellular target of histatin 5 on Candida albicans is the energized mitochondrion.
J. Biol. Chem. 1999, 274, 7286–7291
274. Vylkova, S.; Jang, W.S.; Li, W.; Nayyar, N.; Edgerton, M. Histatin 5 initiates osmotic stress
response in Candida albicans via activation of the Hog1 mitogen-activated protein kinase
pathway. Eukaryot. Cell 2007, 6, 1876–1888.
275. Wagener, J.; Schneider, J.J.; Baxmann, S.; Kalbacher, H.; Borelli, C.; Nuding, S.; Küchler, R.;
Wehkamp, J.; Kaeser, M.D.; Mailänder-Sanchez, D.; et al. A peptide derived from the highly
conserved protein Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is involved in tissue
protection by different antifungal strategies and epithelial immunomodulation. J. Invest.
Dermatol. 2013, 133, 144–153.
276. Aslam, R.; Atindehou, M.; Lavaux, T.; Haï kel, Y.; Schneider, F.; Metz-Boutigue, M.H.
Chromogranin A-derived peptides are involved in innate immunity. Curr. Med. Chem. 2012, 19,
4115–4123.
277. Ochoa, M.T.; Stenger, S.; Sieling, P.A.; Thoma-Uszynski, S.; Sabet, S.; Cho, S.; Krensky, A.M.;
Rollinghoff, M.; Nunes Sarno, E.; Burdick, A.E.; et al. T-cell release of granulysin contributes to
host defense in leprosy. Nat. Med. 2001, 7, 174–179.
278. Stewart, S.E.; Kondos, S.C.; Matthews, A.Y.; D'Angelo, M.E.; Dunstone, M.A.; Whisstock, J.C.;
Trapani, J.A.; Bird, P.I. The Perforin Pore Facilitates the Delivery of Cationic Cargos. J. Biol.
Chem. 2014, 289, 9172–9181.
279. Diamond, G.; Beckloff, N.; Weinberg, A.; Kisich, K.O. The roles of antimicrobial peptides in
innate host defense. Curr. Pharm Des. 2009, 15, 2377–2392.
280. Leung, T.F.; Ching, K.W.; Kong, A.P.; Wong, G.W.; Chan, J.C.; Hon, K.L. Circulating LL-37 is
a biomarker for eczema severity in children. J. Eur. Acad. Dermatol. Venereol. 2012, 26,
518–522.
281. Weber, G.; Chamorro, C.I.; Granath, F.; Liljegren, A.; Zreika, S.; Saidak, Z.; Sandstedt, B.;
Rotstein, S.; Mentaverri, R.; Sánchez, F.; et al. Human antimicrobial protein hCAP18/LL-37
promotes a metastatic phenotype in breast cancer. Breast Cancer Res. 2009, 11, R6.
282. Cheng, W.L.; Wang, C.S.; Huang, Y.H.; Liang, Y.; Lin, P.Y.; Hsueh, C.; Wu, Y.C.; Chen, W.J.;
Yu, C.J.; Lin, S.R.; et al. Overexpression of a secretory leukocyte protease inhibitor in human
gastric cancer. Int. J. Cancer 2008, 123, 1787–1796.
283. Emson, C.L.; Fitzmaurice, S.; Lindwall, G.; Li, K.W.; Hellerstein, M.K.; Maibach, H.I.;
Liao, W.; Turner, S.M. A pilot study demonstrating a non-invasive method for the measurement
of protein turnover in skin disorders: Application to psoriasis. Clin. Transl. Med. 2013, 2, 12.
284. Welling, M.M.; Paulusma-Annema, A.; Balter, H.S.; Pauwels, E.K.; Nibbering, P.H.
Technetium-99m labeled antimicrobial peptides discriminate between bacterial infections and
sterile inflammations. Eur. J. Nucl. Med. 2000, 27, 292–301.
285. Saeed, S.; Zafar, J.; Khan, B.; Akhtar, A.; Qurieshi, S.; Fatima, S.; Ahmad, N.; Irfanullah, J.
Utility of ⁹⁹mTc-labelled antimicrobial peptide ubiquicidin (29–41) in the diagnosis of diabetic
foot infection. Eur. J. Nucl. Med. Mol. Imaging 2013, 40, 737–743.
286. Aryana, K.; Hootkani, A.; Sadeghi, R.; Davoudi, Y.; Naderinasab, M.; Erfani, M.; Ayati, N.
(99m)Tc-labeled ubiquicidin scintigraphy: A promising method in hip prosthesis infection
diagnosis. Nuklearmedizin 2012, 51, 133–139.
287. Wang, G. Antimicrobial Peptides: Discovery, Design and Novel Therapeutic Strategies; CABI:
Wallingford, UK, 2010.
288. Forde, E.; Humphreys, H.; Greene, C.M.; Fitzgerald-Hughes, D.; Devocelle, M. The potential of
host defence peptide prodrugs as neutrophil elastase-dependent anti-infective agents for cystic
fibrosis. Antimicrob. Agents. Chemother. 2013, 58, 978–985.
289. Reynolds, N.L.; de Cecco, M.; Taylor, K.; Stanton, C.; Kilanowski, F.; Kalapothakis, J.; Seo, E.;
Uhrin, D.; Campopiano, D.; Govan, J.; et al. Peptide fragments of a beta-defensin derivative with
potent bactericidal activity. Antimicrob. Agents Chemother. 2010, 54, 1922–1929
290. Roelandts, R. The history of phototherapy: Something new under the sun? J. Am. Acad.
Dermatol. 2002, 46, 926–930.
291. Wang, T.T.; Nestel, F.P.; Bourdeau, V.; Nagai, Y.; Wang, Q.; Liao, J.; Tavera-Mendoza, L.;
Lin, R.; Hanrahan, J.W.; Mader, S.; et al. Cutting edge: 1,25-dihydroxyvitamin D3 is a direct
inducer of antimicrobial peptide gene expression. J. Immunol. 2004, 173, 2909–2912.
292. Gombart, A.F.; Borregaard, N.; Koeffler, H.P. Human cathelicidin antimicrobial peptide
(CAMP) gene is a direct target of the vitamin D receptor and is strongly up-regulated in myeloid
cells by 1,25-dihydroxyvitamin D3. FASEB J. 2005, 19, 1067–1077.
293. Ala-Houhala, M.J.; Karppinen, T.; Vähävihu, K.; Kautiainen, H.; Dombrowski, Y.; Snellman, E.;
Schauber, J.; Reunala, T. Narrow-band Ultraviolet B Treatment Boosts Serum 25-hydroxyvitamin
D in Patients with Psoriasis on Oral Vitamin D Supplementation. Acta Derm. Venereol. 2014, 94,
146–151.
294. Steinmann, J.; Halldórsson, S.; Agerberth, B.; Gudmundsson, G.H. Phenylbutyrate (PBA)
induces antimicrobial peptide expression. Antimicrob. Agents Chemother. 2009, 53, 5127–5133.
295. Gan, Y.; Cui, X.; Ma, T.; Liu, Y.; Li, A.; Huang, M. Paeoniflorin Upregulates β-Defensin-2
Expression in Human Bronchial Epithelial Cell Through the p38 MAPK, ERK, and NF-κB
Signaling Pathways. Inflammation. 2014, in press.
296. Kim, B.J.; Rho, Y.K.; Lee, H.I.; Jeong, M.S.; Li, K.; Seo, S.J.; Kim, M.N.; Hong, C.K. The
effect of calcipotriol on the expression of human beta defensin-2 and LL-37 in cultured human
keratinocytes. Clin. Dev. Immunol. 2009, 2009, 645898.
297. Sunkara, L.T.; Zeng, X.; Curtis, A.R.; Zhang, G. Cyclic AMP synergizes with butyrate in
promoting β-defensin 9 expression in chickens. Mol. Immunol. 2014, 57, 171–180.
298. Cederlund, A.; Nylén, F.; Miraglia, E.; Bergman, P.; Gudmundsson, G.H.; Agerberth, B. Label-
Free Quantitative Mass Spectrometry Reveals Novel Pathways Involved in LL-37 Expression. J.
Innate Immun. 2014, 6, 365–376.
299. Liu, Q;, Liu, J.; Roschmann, K.I.; van Egmond, D.; Golebski, K.; Fokkens, W.J.; Wang, D.; van
Drunen, C.M. Histone deacetylase inhibitors up-regulate LL-37 expression independent of
toll-like receptor mediated signalling in airway epithelial cells. J. Inflamm. (Lond). 2013, 10, 15.
300. Strandberg, K.L.; Richards, S.M.; Gunn, J.S. Cathelicidin antimicrobial peptide expression is not
induced or required for bacterial clearance during salmonella enterica infection of human
monocyte-derived macrophages. Infect. Immun. 2012, 80, 3930–3938.
301. Yin, Q.; Xu, X.; Lin, Y.; Lv, J.; Zhao, L.; He, R. Ultraviolet B irradiation induces skin
accumulation of plasmacytoid dendritic cells: A possible role for chemerin. Autoimmunity. 2013,
47, 185–192.
302. Karlsson, J.; Carlsson, G.; Larne, O.; Andersson, M.; Pütsep, K. Vitamin D3 induces pro-LL-37
expression in myeloid precursors from patients with severe congenital neutropenia. J. Leukoc.
Biol. 2008, 84, 1279–1286.
303. Martineau, A.R.; Wilkinson, K.A.; Newton, S.M.; Floto, R.A.; Norman, A.W.; Skolimowska, K.;
Davidson, R.N.; Sørensen, O.E.; Kampmann, B.; Griffiths, C.J.; et al. IFN-gamma- and
TNF-independent vitamin D-inducible human suppression of mycobacteria: The role of
cathelicidin LL-37. J. Immunol. 2007, 178, 7190–7198.
304. Cederlund, A.; Kai-Larsen, Y.; Printz, G.; Yoshio, H.; Alvelius, G.; Lagercrantz, H.;
Strömberg, R.; Jörnvall, H.; Gudmundsson, G.H.; Agerberth, B. Lactose in human breast milk an
inducer of innate immunity with implications for a role in intestinal homeostasis. PLoS One
2013, 8, e53876.
305. Jiang, W.; Sunkara, L.T.; Zeng, X.; Deng, Z.; Myers, S.M.; Zhang, G. Differential regulation of
human cathelicidin LL-37 by free fatty acids and their analogs. Peptides 2013, 50C, 129–138.
306. Zeng, X.; Sunkara, L.T.; Jiang, W.; Bible, M.; Carter, S.; Ma, X.; Qiao, S.; Zhang, G. Induction
of porcine host defense peptide gene expression by short-chain fatty acids and their analogs.
PLoS One 2013, 8, e72922.
307. Sherman, H.; Chapnik, N.; Froy, O. Albumin and amino acids upregulate the expression of
human beta-defensin 1. Mol. Immunol. 2006, 43, 1617–1623.
308. Fehlbaum, P.; Rao, M.; Zasloff, M.; Anderson, G.M. An essential amino acid induces epithelial
beta-defensin expression. Proc. Natl. Acad. Sci. USA 2000, 97, 12723–12728.
309. Rivas-Santiago, C.E.; Rivas-Santiago, B.; León, D.A.; Castañeda-Delgado, J.; Hernández Pando, R.
Induction of β-defensins by l-isoleucine as novel immunotherapy in experimental murine
tuberculosis. Clin. Exp. Immunol. 2011, 164, 80–89.
310. Pernet, I.; Reymermier, C.; Guezennec, A.; Branka, J.E.; Guesnet, J.; Perrier, E.;
Dezutter-Dambuyant, C.; Schmitt, D.; Viac, J. Calcium triggers beta-defensin (hBD-2 and
hBD-3) and chemokine macrophage inflammatory protein-3 alpha (MIP-3alpha/CCL20)
expression in monolayers of activated human keratinocytes. Exp. Dermatol. 2003, 12, 755–760.
311. Talukder, P.; Satho, T.; Irie, K.; Sharmin, T.; Hamady, D.; Nakashima, Y.; Kashige, N.;
Miake, F. Trace metal zinc stimulates secretion of antimicrobial peptide LL-37 from Caco-2 cells
through ERK and p38 MAP kinase. Int. Immunopharmacol. 2011, 11, 141–144.
312. Schauber, J.; Iffland, K.; Frisch, S.; Kudlich, T.; Schmausser, B.; Eck, M.; Menzel, T.;
Gostner, A.; Lührs, H.; Scheppach, W. Histone-deacetylase inhibitors induce the cathelicidin
LL-37 in gastrointestinal cells. Mol. Immunol. 2004, 41, 847–854.
313. Thivierge, K.; Cotton, S.; Schaefer, D.A.; Riggs, M.W.; To, J.; Lund, M.E.; Robinson, M.W.;
Dalton, J.P.; Donnelly, S.M. Cathelicidin-like helminth defence molecules (HDMs): Absence of
cytotoxic, anti-microbial and anti-protozoan activities imply a specific adaptation to immune
modulation. PLoS Negl. Trop. Dis. 2013, 7, e2307.
314. Yeaman, M.R.; Yount, N.Y. Mechanisms of antimicrobial peptide action and resistance.
Pharmacol. Rev. 2003, 55, 27–55.
315. Peschel, A. How do bacteria resist human antimicrobial peptides? Trends Microbiol. 2002, 10,
179–186.
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Pharmaceuticals 2014, 7, 545-594; doi:10.

Human Antimicrobial Peptides and Proteins

Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical

Keywords: antimicrobial chemokines; antimicrobial neuropeptides; antimicrobial proteins;

Table 1. Discovery timeline of select human antimicrobial peptides and proteins 1.

2. Identification of Human Antimicrobial Peptides

2.1. Human Defensins

2.2. Human Histatins: Two Genes Multiple Peptides

2.3. Human Cathelicidins: One Gene Multiple Peptides

2.4. Human Dermcidin

2.5. Human Hepcidins

2.6. Human AMPs Derived from Known Proteins

2.7. Antimicrobial Chemokines and AMPs from Human Immune Cells

2.8. Antimicrobial Neuropeptides

2.9. Beta-Amyloid Peptides

2.10. Human Antimicrobial Proteins

3. Antimicrobial and Anticancer Activities of Human Antimicrobial Peptides

3.1. Antibacterial Activities

3.2. Antiviral Activity

3.3. Antifungal Activity

3.4. Antiparasitic Activity

3.5. Anticancer Activity

3.6. Cytotoxic Effects of Human AMPs

3.7. Other Biological Functions of Human AMPs

4. Three-Dimensional Structures of Human Antimicrobial Peptides

Table 2. Properties of selected human antimicrobial peptides with known 3D structure 1.

4.1. The α-Helical Family: Histatins, Cathelicidins, Dermcidin, and Granulysin

Human cathelicidin LL-37. To understand the structural basis of antimicrobial activity, 3D

Figure 1. Three-dimensional structures of human antimicrobial peptides from the α-helical

4.2. The β Family: α-Defensins

4.3. The αβ Family: β-Defensins, Antimicrobial Chemokines, RNases, and RegIIIα

5. Mechanism of Action of Human Antimicrobial Peptides

5.1. Targeting Bacterial Cell Wall

5.2. Targeting Bacterial Inner Membranes

5.3. Cell-Penetrating Peptides and Intracellular Targets

6. Concluding Remarks and Potential Therapeutic Strategies

Table 3. Select human antimicrobial peptides and their proposed targets.

Table 4. Some known factors that induce antimicrobial peptide expression

The author does not declare a conflict of interest.

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