Structural and Functional Analysis Reveals the Catalytic Mechanism and Substrate Binding Mode of the Broad-spectrum Endolysin Ply2741

Overview

Journal Virulence

Date 2025 Jan 15

PMID 39810299

Authors

Shuang Wang

Xinxin Li

Jiahui Ma

Xiaochao Duan

Haiyan Wang

Linkang Wang

Dayue Hu

Wenwu Jiang

Xiangmin Li

Ping Qian

Affiliations

Soon will be listed here.

Abstract

The emergence of antibiotic-resistant bacteria has attracted interest in the field of endolysins. Here, we analyzed the diversity of endolysins and identified a new endolysin, Ply2741, that exhibited broad-spectrum bactericidal activity. Our results demonstrated that Ply2741 could effectively eradicate multidrug-resistant gram-positive pathogens and . Structural analysis revealed that the bactericidal activity of Ply2741 depends on the classic "Cys-His-Asn" catalytic triad. Site-directed mutagenesis results further identified that the conserved residue Gln29, located near the catalytic triad, also contributes to the lytic activity of Ply2741. Furthermore, the key residues (R189 and W250) in the Ply2741 cell wall binding domain (CBD) responsible for binding to peptidoglycan were revealed by molecular docking and fluorescence-activated cell sorting (FACS) analysis. Ply2741 demonstrates a broad lytic spectrum, with significant bactericidal activity against , s, and species. To the best of our knowledge, we found that residue Gln29 participated in the lytic activity of endolysin for the first time. Additionally, we systematically elucidate the binding mode and key residues of the Ply2741CBD. This study proposes Ply2741 as a potential antibiotic substitute and provides a structural basis for the modification and design of endolysins.

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