ANTIMICROBIAL

PEPTIDES
Their History, Evolution, and Functional Promiscuity
1.1 Introduction: The History of
Microbial Peptides
• 1939  PROKARYOTIC -
Gramicidins = Activity against a
wide range of Gram + bacteria.

• First AMP’s commercially

manufactured as antibiotics
Bacillus brevis

Why plants & insects, which lack an adaptive immune

system, also remain free from infections for most the time?
 EUKARYOTES also produce AMP’S

• 1896  Substance lethal to bread yeast was present in

wheat flour.
 1920  Lysosyme
 1928  Penicillin
 1940  Therapeutic use
 1945  Nobel Prize for
Medicine

GOLDEN AGE OF
Alexander Fleming
ANTIBIOTICS
 Multidrug resistant Microbial Pathogens
1960’s
• True origin of research
• Cationic proteins  Neutrophils = kill bacteria via oxygen-
independent mechanisms

• 1962  ANIMALS – Bombinin

 Lactoferrin (Milk)

 70’s & 80’s  LEUKOCYTES

alpha-defensins

Bombina variegate
• 1981 (Boman et al.) CECROPINS 
Cationic – Bacteria into pupae of the
silk moth

Hyalophora cecropia

 1987 (Zasloff et al.) MAGAININS

 Cationic AMP’s

Xenopus laevis
 • 90’s  Beta & Theta defensins

 90’s  LYSOZYME –
Antimicrobial activity
involving NON-ENZYMATIC
mechanisms

Bovine granulocytes

AMP’s may play a role in the defense systems of organisms

lacking an adaptive immune system
AMP’s may play a role in the defense systems of organisms
lacking an adaptive immune system

 The DELETION of a gene encoding an AMP rendered the

insect susceptible to a MASSIVE FUNGAL INFECTION

Drosophila melanogaster
Highlights
 Identified in sites exposed to microbes
 Their production could be constitutive or induced
 Not exclusive
 Each tissue has its own spectrum of AMP’s

 Minimal inhibitory concentrations for antimicrobial activity in

vitro are higher than in vivo
 Accumulate
 Act synergistically
 Structurally similar and same host
 Structurally dissimilar and different host
Highlights
 As more has been learnt about AMP’s it has become
somewhat arbitrary as to their precise definition.
 Ubiquitous
 Non-specific mode of antimicrobial action
 *Some do not appear to exert direct antimicrobial activity

 2000 AMP’s  Databases

 Allowed comparisons: STRUCTURE, FUNCTION,
MECHANISMS.
 Ancient origins
 1.2 AMPs: Evolutionarily Ancient
Molecules
• Most of current understanding of AMP’s  Amphibian skin secretions
• 10-20 AMP’s
• DNA, protein  sequence data

Ranidae Hylidae
RANAEs (Asia, North America, Europe) HASs (South America)
HAs (Australia)
  Precursor proteins are highly
conserved
 Genes arose from common ancestral
locus. Diversified by duplication and
divergence of loci.
 Diversity  Random substitutions in
DNA

Represents the successful EVOLUTION of a

DEFENSE SYSTEM that maximizes host
protection against RAPIDLY CHANGING
MICROBIAL BIOTA
 •Defensins Vertebrates

 Alpha & beta  triple-stranded antiparallel beta-sheet

configurations
 Theta  cyclized product of a head-to-tail ligation of two
truncated alpha-defensins
 All from a single precursor
 Theta-defensins  rhesus macaque monkeys and
baboons
 Premature stop codon
 Pseudogene (aminoglycosides) = RETROCYCLINS
 Potent antiviral activity (HIV)
 Duplicated genes  NOVEL (or more) SPECIALIZED
FUNCTION

 Inhibit the activity & synthesis of

alpha-amylase
Plants

 Beta-defensins = venom

 Non venom tissues ???

 EVOLUTIONARY ADVANTAGE
Platypus
1.3 Multifunctional Molecules
• Alpha-MSH
• Control inflammation  generation of anti-inflammatory cytokines
• Widespread distribution
• Barrier cells & immune cells
• Membranolytic action against bacteria

Functional promiscuity
Defensins vs. Chemokines
AMP’s  variety of biological roles
• Immunomodulatory functions
• Direct antimicrobial activity
1.3.1 Defensins as Effectors of
Immunity
• Human beta-defensins  Epithelial tissues
• Direct action Gram+ Gram-, fungi, viruses and parasites
• Regulating inflammation
• Healing inflamed tissue
• Function as chemoattractant for immune cells
 HBDs can serve as modulators of both the innate and adaptive
immune response to infection
 HBDs directly
promote a
proinflammatory
immune response
by binding to
various cell
receptors.

 HBDs levels
decrease as the
microbial treat is
nullified 
Suppression of
proinflammatory
activity.
 1.3.2 Defensins and Wound Healing
• HBD 1-4  SKIN Healthy and burned
• Migration and proliferation of epidermal keratinocytes
• WOUND HEALING

• HBD2  acute/chronic wounds (ocular surface, mucosal barrier in the

intestine)
• High –glucose  reduces HBD2 expression
1.3.3 Defensins and Canine Coat
Color  CBD103 – Canine beta-
defensing
 Potent antibacterial – skin of
dogs
 Determines color of dog coats
 Molecular characterization of Antimicrobial
Peptide Genes of the Carpenter Ant
Camponotus floridanus
 Cystein rich: defensin 1, defensing 2
 Glycine rich: hymenoptaecin
 Lack of pleural gland
 Antimicrobial compunds
 Obligate intracellular
endosymbiont
 Tolerated by host’s defense
mechanisms

 Gene is long and encodes a multipeptide precursor, the

proteolytic processing of which possibly leads to a massive
amplification of the antimicrobial response.
From antimicrobial to anticancer peptides. A
review

From antimicrobial to anticancer peptides. A review

Diana Gaspar, A. Salomé Veiga, Miguel A. R. B. Castanho
Front Microbiol. 2013; 4: 294. Published online 2013 Oct 1. doi: 10.3389/fmicb.2013.00294
ANTIMICROBIAL
PEPTIDES
Their History, Evolution, and Functional Promiscuity