Abstract
Cationic antimicrobial peptides and their therapeutic potential have garnered growing interest because of the proliferation of bacterial resistance. However, the discovery of new antimicrobial peptides from animals has proven challenging due to the limitations associated with conventional biochemical purification and difficulties in predicting active peptides from genomic sequences, if known. As an example, no antimicrobial peptides have been identified from the American alligator, Alligator mississippiensis, although their serum is antimicrobial. We have developed a novel approach for the discovery of new antimicrobial peptides from these animals, one that capitalizes on their fundamental and conserved physico-chemical properties. This sample-agnostic process employs custom-made functionalized hydrogel microparticles to harvest cationic peptides from biological samples, followed by de novo sequencing of captured peptides, eliminating the need to isolate individual peptides. After evaluation of the peptide sequences using a combination of rational and web-based bioinformatic analyses, forty-five potential antimicrobial peptides were identified, and eight of these peptides were selected to be chemically synthesized and evaluated. The successful identification of multiple novel peptides, exhibiting antibacterial properties, from Alligator mississippiensis plasma demonstrates the potential of this innovative discovery process in identifying potential new host defense peptides.
Highlights
There has been a growing interest in cationic antimicrobial peptides (CAMPs) as a potential source of new therapeutics with which to address the growing problem of bacterial antibioticPLOS ONE | DOI:10.1371/journal.pone.0117394 February 11, 2015Bioprospecting American Alligator Host Defense resistance [1, 2]
The first step in the bioprospecting process used to identify novel alligator CAMPs employed hydrogel microparticles based on cross-linked N-isopropylacrylamide copolymer frameworks, which are central to the CAMP discovery process [8, 9]
In order to overcome the limitations associated with current approaches to CAMP discovery, we have developed a novel and promising method for identifying new and potentially useful antimicrobial peptides
Summary
The American alligator (Alligator mississippiensis) and other crocodilians are evolutionarily ancient animals whose plasma and leukocyte extracts have been shown to exhibit antimicrobial activity [3,4,5]. This antimicrobial potency may be attributable at least in part to the presence of CAMPs in the alligator plasma and extracts. Despite the interest in the American alligator and the antimicrobial peptides that they may produce, no CAMPs have been identified from their blood or tissues to date [3, 6]
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