title: Bacillus subtilis CwlP of the SP-{beta} prophage has two novel peptidoglycan hydrolase domains, muramidase and cross-linkage digesting DD-endopeptidase.
abstract: For bacteria and bacteriophages, cell wall digestion by hydrolases is a very important event. We investigated one of the proteins involved in cell wall digestion, the yomI gene product (renamed CwlP). The gene is located in the SP-β prophage region of the Bacillus subtilis chromosome. Inspection of the Pfam database indicates that CwlP contains soluble lytic transglycosylase (SLT) and peptidase M23 domains, which are similar to Escherichia coli lytic transglycosylase Slt70, and the Staphylococcus aureus Gly-Gly endopeptidase LytM, respectively. The SLT domain of CwlP exhibits hydrolytic activity toward the B. subtilis cell wall; however, reverse phase (RP)-HPLC and mass spectrometry revealed that the CwlP-SLT domain has only muramidase activity. In addition, the peptidase M23 domain of CwlP exhibited hydrolytic activity and could cleave d-Ala-diaminopimelic acid cross-linkage, a property associated with dd-endopeptidases. Remarkably, the M23 domain of CwlP possessed a unique Zn(2+)-independent endopeptidase activity; this contrasts with all other characterized M23 peptidases (and enzymes similar to CwlP), which are Zn(2+) dependent. Both domains of CwlP could hydrolyze the peptidoglycan and cell wall of B. subtilis. However, the M23 domain digested neither the peptidoglycans nor the cell walls of S. aureus or Streptococcus thermophilus. The effect of defined point mutations in conserved amino acid residues of CwlP is also determined.
authors.affiliation: Interdisciplinary Graduate School of Science and Technology, Department of Life Sciences, Research Center for Human and Environmental Sciences, Shinshu University, Nagano 386-8567, Japan.
authors.forename: I Putu
authors.initials: IP
authors.lastname: Sudiarta
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Fukushima
authors.forename: Junichi
authors.initials: J
authors.lastname: Sekiguchi
mesh_terms: D001412:Bacillus subtilis / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics*; D002412:Cations; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D010450:Endopeptidases / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D009113:Muramidase / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D010457:Peptidoglycan / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D011108:Polymers / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D039002:Prophages; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D015032:Zinc / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D002412:Cations; D010457:Peptidoglycan; D011108:Polymers; D011134:Polysaccharides; D009113:Muramidase; C019637:DD-endopeptidase; D010450:Endopeptidases; D015032:Zinc