Biochemical Investigation of the Role of Loop 1 Residues in LytM
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.
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BibTeX
@misc{gonzalez2026biochemical,
title = {{Biochemical Investigation of the Role of Loop 1 Residues in LytM}},
author = {Sophia Y. Gonzalez and Cassidy A. McIntyre and Amber L. B. Mogg and Manya P. Sharma and Sierra J. Williams},
howpublished = {ChemRxiv},
year = {2026},
month = aug,
doi = {10.26434/chemrxiv.15008046/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15008046/v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Biochemical Investigation of the Role of Loop 1 Residues in LytM
AU - Sophia Y. Gonzalez
AU - Cassidy A. McIntyre
AU - Amber L. B. Mogg
AU - Manya P. Sharma
AU - Sierra J. Williams
PY - 2026
DA - 2026/08/30
PB - ChemRxiv
DO - 10.26434/chemrxiv.15008046/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15008046/v1
AB - 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.
ER -