Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 Strains

In our previous work, we showed that short antimicrobial hexapeptides (AMPs) containing three Trp and three Arg residues had a potent antibiofilm activity against a pathogenic Gram-positive <i>Staphylococcus aureus</i> MRSA strain. However, the activity of these hexapeptides against a Gr...

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Main Authors: Gopal Ramamourthy, Hiroaki Ishida, Hans J. Vogel
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/4/826
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author Gopal Ramamourthy
Hiroaki Ishida
Hans J. Vogel
author_facet Gopal Ramamourthy
Hiroaki Ishida
Hans J. Vogel
author_sort Gopal Ramamourthy
collection DOAJ
description In our previous work, we showed that short antimicrobial hexapeptides (AMPs) containing three Trp and three Arg residues had a potent antibiofilm activity against a pathogenic Gram-positive <i>Staphylococcus aureus</i> MRSA strain. However, the activity of these hexapeptides against a Gram-negative <i>Pseudomonas aeruginosa</i> PA01 strain was relatively poor. Herein, we tested the longer 13-residue synthetic AMP tritrpticin-NH<sub>2</sub> (Tritrp) and several of its analogs as potential antibiofilm agents that can prevent biofilm formation (MBIC) and/or cause biofilm dissolution (MBEC) for two <i>P. aeruginosa</i> PA01 strains, one of which expressed the GFP protein. Tritrp, a porcine cathelicidin, is currently the only known naturally occurring cationic AMP that has three Trp in sequence (WWW), a feature that was found to be important in our previous study. Our results show that several Tritrp analogs were effective. In particular, analogs with Pro substitutions that had altered peptide backbone structures compared to the naturally occurring amphipathic two-turn structure showed more potent MBIC and MBEC antibiofilm activities. Selectivity of the peptides towards <i>P. aeruginosa</i> could be improved by introducing the non-proteinogenic amino acid 2,3-diaminopropionic acid, rather than Arg or Lys, as the positively charged residues. Using <sup>1</sup>H NMR spectroscopy, we also reinvestigated the role of the two Pro residues in cis–trans isomerism of the peptide in aqueous solution. Overall, our results show that the WWW motif embedded in longer cationic AMPs has considerable potential to combat biofilm formation in pathogenic Gram-negative strains.
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spelling doaj-art-ea14f5ebb71b438d8d5658a146c5d6032025-08-20T02:04:06ZengMDPI AGMolecules1420-30492025-02-0130482610.3390/molecules30040826Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 StrainsGopal Ramamourthy0Hiroaki Ishida1Hans J. Vogel2Biochemistry Research Group, Department of Biological Sciences, University of Calgary, Calgary, AB T2N 1N4, CanadaBiochemistry Research Group, Department of Biological Sciences, University of Calgary, Calgary, AB T2N 1N4, CanadaBiochemistry Research Group, Department of Biological Sciences, University of Calgary, Calgary, AB T2N 1N4, CanadaIn our previous work, we showed that short antimicrobial hexapeptides (AMPs) containing three Trp and three Arg residues had a potent antibiofilm activity against a pathogenic Gram-positive <i>Staphylococcus aureus</i> MRSA strain. However, the activity of these hexapeptides against a Gram-negative <i>Pseudomonas aeruginosa</i> PA01 strain was relatively poor. Herein, we tested the longer 13-residue synthetic AMP tritrpticin-NH<sub>2</sub> (Tritrp) and several of its analogs as potential antibiofilm agents that can prevent biofilm formation (MBIC) and/or cause biofilm dissolution (MBEC) for two <i>P. aeruginosa</i> PA01 strains, one of which expressed the GFP protein. Tritrp, a porcine cathelicidin, is currently the only known naturally occurring cationic AMP that has three Trp in sequence (WWW), a feature that was found to be important in our previous study. Our results show that several Tritrp analogs were effective. In particular, analogs with Pro substitutions that had altered peptide backbone structures compared to the naturally occurring amphipathic two-turn structure showed more potent MBIC and MBEC antibiofilm activities. Selectivity of the peptides towards <i>P. aeruginosa</i> could be improved by introducing the non-proteinogenic amino acid 2,3-diaminopropionic acid, rather than Arg or Lys, as the positively charged residues. Using <sup>1</sup>H NMR spectroscopy, we also reinvestigated the role of the two Pro residues in cis–trans isomerism of the peptide in aqueous solution. Overall, our results show that the WWW motif embedded in longer cationic AMPs has considerable potential to combat biofilm formation in pathogenic Gram-negative strains.https://www.mdpi.com/1420-3049/30/4/826antibiofilm peptidesantimicrobial peptidesbiofilmsdiaminopropionic acid<i>Pseudomonas aeruginosa</i>tryptophan
spellingShingle Gopal Ramamourthy
Hiroaki Ishida
Hans J. Vogel
Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 Strains
Molecules
antibiofilm peptides
antimicrobial peptides
biofilms
diaminopropionic acid
<i>Pseudomonas aeruginosa</i>
tryptophan
title Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 Strains
title_full Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 Strains
title_fullStr Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 Strains
title_full_unstemmed Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 Strains
title_short Antibiofilm Activities of Tritrpticin Analogs Against Pathogenic <i>Pseudomonas aeruginosa</i> PA01 Strains
title_sort antibiofilm activities of tritrpticin analogs against pathogenic i pseudomonas aeruginosa i pa01 strains
topic antibiofilm peptides
antimicrobial peptides
biofilms
diaminopropionic acid
<i>Pseudomonas aeruginosa</i>
tryptophan
url https://www.mdpi.com/1420-3049/30/4/826
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AT hansjvogel antibiofilmactivitiesoftritrpticinanalogsagainstpathogenicipseudomonasaeruginosaipa01strains