Theoretical Design via Umbrella Sampling and Experimental Investigation of a New Mutation in Cap37 to Increase its Antibacterial Property

Hedayati, Mehri; Mahnam, Karim; Saffar, Behnaz

Theoretical Design via Umbrella Sampling and Experimental Investigation of a New Mutation in Cap37 to Increase its Antibacterial Property

Authors

Mehri Hedayati ¹

Karim mahnam ¹

Behnaz Saffar ²

¹ shahrekord University

² University of shshrekord

Abstract

Abstract

The increase in antibacterial resistance due to the high consumption of antibiotics is a looming crisis for humanity. Natural antibacterial compounds, such as cationic antibacterial peptides, hold a significant position, as the likelihood of developing resistance against them is low. CAP37, or AZU1, is a protein derived from the granules of human neutrophils and functions as a natural antibiotic. The residues 20-44 of CAP37 exhibit antibacterial activity. In this study, two mutations were designed in the native CAP37 peptide to enhance its antibacterial activity. Molecular dynamics simulations of the peptides in water were conducted for 50 ns, along with umbrella sampling MD simulations to obtain reliable potential of mean force (PMF) profiles for each peptide's passage through lipid A, a component of the bilayer membrane. The results indicated that the peptide containing the SWRW sequence exhibited lower aggregation properties in water and a greater tendency to traverse the lipid A bilayer membrane compared to the native (SQRS) or WWRS sequences. Therefore, it can be concluded that CAP37 with the SWRW sequence has enhanced antimicrobial activity. To validate the theoretical findings, both the natural and SWRW mutant peptides were synthesized. The minimum inhibitory concentration (MIC) test conducted on various Gram-negative and Gram-positive bacteria, including Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus subtilis, demonstrated that the mutated peptide exhibited antibacterial activity, thereby confirming the theoretical results.

Keywords

Antimicrobial pepide

Cap37 protein

Molecular dynamic simulation

MIC test

Mutated peptide

Subjects

Pharmaceutical Biotechnology

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