Characterization of a chromosomally encoded glycylglycine endopeptidase of Staphylococcus aureus.
The authors previously reported the cloning of a lytic-enzyme-encoding gene, lytM, from an autolysis-defective mutant of Staphylococcus aureus. In the present work, the lytM gene was overexpressed in Escherichia coli and the product was purified to homogeneity by affinity chromatography and HPLC. Biochemical analysis of LytM-cleaved peptidoglycan fragments indicated that LytM is a glycylglycine endopeptidase. Immunoelectron microscopic studies with anti-LytM rabbit IgG showed that LytM is expressed during the early exponential phase and is overexpressed in an autolysis-defective mutant compared with the parent strain. Also, a uniform distribution of gold particles on the surface of actively growing bacterial cells indicates that LytM plays a role in cell growth. Northern blot analyses of lytM expression in two global regulatory mutants, agr and sar, showed that expression of lytM is increased about twofold in these mutants as compared with the parents. Protein homology searches revealed that LytM could be a member of the zinc protease family, as it contained a homologous 38-amino-acid motif, Tyr-X-His-X11-Val-X12/20-Gly-X5-6-His. Atomic absorption spectrometric analysis of LytM revealed the presence of 0.9 mol zinc (mol LytM)(-1).
Ramadurai
Lakshmi
L
Lockwood
Katherine J
KJ
Lockwood
J
J
Nadakavukaren
Mathew J
MJ
Jayaswal
Radheshyam K
RK
D000818:Animals; D001426:Bacterial Proteins; D001433:Bacteriolysis; D015153:Blotting, Western; D002876:Chromosomes, Bacterial / Q000235:genetics; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D008024:Ligands; D008854:Microscopy, Electron; D011817:Rabbits; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D013379:Substrate Specificity; D014158:Transcription, Genetic; D015032:Zinc / Q000032:analysis
D001426:Bacterial Proteins; D008024:Ligands; D010450:Endopeptidases; C120985:LytM protein, Staphylococcus aureus; D015032:Zinc