Year: 2026
Type: preprint
Abstract: Infections caused by drug-resistant bacteria impact the lives of millions across the globe. To combat these infections, peptidoglycan hydrolases (PGHs) are gaining interest due to their ability to effectively kill these pathogens with low rates of evoking resistance pathways. One such PGH includes LytM, which is an autolysin produced by Staphylococcus aureus. While an engineered variant of this enzyme has shown antimicrobial activity, its mechanism is poorly understood. The loop 1 region was proposed to be responsible for a new cleavage site that has not been reported for other M23 enzymes. However, it is unclear what residues in loop 1 are key for catalytic activity. In this work, a biochemical analysis was used to identify sites that were important for activity. An alanine scan of the loop 1 region of the enzyme revealed key hotspots for hydrolysis. We then used site-saturation mutagenesis to identify the structure-function relationship of these sites. The function of these residues was then tested in a homologous enzyme, lysostaphin, to determine how well activity is conserved. Collectively, this work confirms residues that are important for activity of LytM and related enzymes.
Source: ChemRxiv
Institutions: Scripps College, Claremont Colleges
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7204827170