Sequence-Activity Relationships and Integrated In Silico and In Vitro Mechanistic Study of the Antimicrobial Peptide PEP-4 against Staphylococcus aureus
- 1Universidade Federal de Pelotas , , Campus Universitário Capão do Leão , , ,
- 2Universidade Federal de Pelotas , , 96010-610 , ,
Abstract
The rise of antimicrobial resistance among ESKAPE pathogens demands novel therapeutic strategies. Here, we evaluate a panel of five synthetic peptides through integrated in vitro and in silico analyses, highlighting PEP-4 as a promising antibacterial lead peptide. Our findings demonstrate the key role of C-terminal hydrophobic patterning in antimicrobial efficacy. PEP-4 exhibits strong antibacterial activity (MIC 15.6–62.5 μg mL–1, corresponding to ∼10.5–41.9 μM) and consistently shows bactericidal effects against both Gram-positive and Gram-negative strains. Molecular dynamics simulations show sustained, charge-selective interaction of PEP-4 with an anionic S. aureus-like bilayer and only intermittent association with a zwitterionic mammalian model, supporting electrostatically driven membrane recognition as the initial step of its mechanism; no bilayer destabilization was observed in either system over the simulated time scale. Additionally, molecular docking suggests potential secondary intracellular protein targets. Notably, PEP-4 retains activity in the presence of lysyl-phospholipid-mediated resistance in Staphylococcus aureus, yet exhibits only transient interactions with zwitterionic mammalian membranes. This selective membrane binding is consistent with low cytotoxicity to humans, corroborated by other in silico predictions. It suggests a potential therapeutic window, pending experimental validation, supporting PEP-4 as a promising peptide scaffold for next-generation antimicrobial design.
Related
- Community-Acquired Staphylococcus argenteus Sequence Type 2250 Bone and Joint Infection, France, 2017
- Staphylococcus aureus in Bovine Mastitis: Pathogenesis, Antimicrobial Resistance, and Emerging Control Strategies
- Machine learning–driven prediction of antimicrobial resistance in Staphylococcus aureus using genomic and allelic signatures
- Staphylococcus pettenkoferi : an underrecognized species with clinically relevant antimicrobial resistance. A comprehensive genomic European study
- Staphylococcus aureus in bovine mastitis in northwestern Iran: patterns of antimicrobial and multidrug resistance, biofilm formation, and virulence/resistance gene carriage
BibTeX
@article{pereira2026sequenceactivity,
title = {{Sequence-Activity Relationships and Integrated In Silico and In Vitro Mechanistic Study of the Antimicrobial Peptide PEP-4 against Staphylococcus aureus}},
author = {Isabel Ladeira Pereira and Isadora Leitzke Guidotti and Danillo de Oliveira Della Senta and Isadora Cosenza Vieira da Silva and Fabiana Kommling Seixas and Leandro Farias Rodrigues and Luciano da Silva Pinto and Daiane Hartwig and Frederico Schmitt Kremer},
journal = {ACS Omega},
year = {2026},
month = oct,
doi = {10.1021/acsomega.6c03955},
url = {https://pubs.acs.org/acsodf/article/11/42/64538/5452490/Sequence-Activity-Relationships-and-Integrated-In},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Sequence-Activity Relationships and Integrated In Silico and In Vitro Mechanistic Study of the Antimicrobial Peptide PEP-4 against Staphylococcus aureus
AU - Isabel Ladeira Pereira
AU - Isadora Leitzke Guidotti
AU - Danillo de Oliveira Della Senta
AU - Isadora Cosenza Vieira da Silva
AU - Fabiana Kommling Seixas
AU - Leandro Farias Rodrigues
AU - Luciano da Silva Pinto
AU - Daiane Hartwig
AU - Frederico Schmitt Kremer
PY - 2026
DA - 2026/10/16
JO - ACS Omega
DO - 10.1021/acsomega.6c03955
UR - https://pubs.acs.org/acsodf/article/11/42/64538/5452490/Sequence-Activity-Relationships-and-Integrated-In
AB - The rise of antimicrobial resistance among ESKAPE pathogens demands novel therapeutic strategies. Here, we evaluate a panel of five synthetic peptides through integrated in vitro and in silico analyses, highlighting PEP-4 as a promising antibacterial lead peptide. Our findings demonstrate the key role of C-terminal hydrophobic patterning in antimicrobial efficacy. PEP-4 exhibits strong antibacterial activity (MIC 15.6–62.5 μg mL–1, corresponding to ∼10.5–41.9 μM) and consistently shows bactericidal effects against both Gram-positive and Gram-negative strains. Molecular dynamics simulations show sustained, charge-selective interaction of PEP-4 with an anionic S. aureus-like bilayer and only intermittent association with a zwitterionic mammalian model, supporting electrostatically driven membrane recognition as the initial step of its mechanism; no bilayer destabilization was observed in either system over the simulated time scale. Additionally, molecular docking suggests potential secondary intracellular protein targets. Notably, PEP-4 retains activity in the presence of lysyl-phospholipid-mediated resistance in Staphylococcus aureus, yet exhibits only transient interactions with zwitterionic mammalian membranes. This selective membrane binding is consistent with low cytotoxicity to humans, corroborated by other in silico predictions. It suggests a potential therapeutic window, pending experimental validation, supporting PEP-4 as a promising peptide scaffold for next-generation antimicrobial design.
SN - 2470-1343
ER -