title: Processing of the lactococcal extracellular serine proteinase.
abstract: Activity of the lactococcal cell envelope-located serine proteinase depends on the presence of membrane-associated lipoprotein PrtM. To differentiate between the action of the proteinase and the action of PrtM in the process of proteinase maturation, an inactive form of the lactococcal proteinase was constructed. This was done by mutating one of the three amino acids thought to constitute the active site of the enzyme. The secreted form of this inactivated proteinase was the same size as the inactive secreted form of the proteinase produced in the absence of PrtM. Both inactive proteinases are larger than the active proteinase. Isolation of proteinase by washing lactococcal cells carrying the complete proteinase gene in a Ca(2+)-free buffer was prevented by the absence of prtM or the absence of a functional active site. We propose that PrtM, during or after membrane translocation of the proteinase, effects the autoproteolytic removal of the N-terminal pro region of the proteinase. Subsequent C-terminal autodigestion results in the release of the enzyme from the lactococcal cells.
authors.affiliation: Department of Genetics, University of Groningen, Haren, The Netherlands.
authors.forename: A J
authors.initials: AJ
authors.lastname: Haandrikman
authors.forename: R
authors.initials: R
authors.lastname: Meesters
authors.forename: H
authors.initials: H
authors.lastname: Laan
authors.forename: W N
authors.initials: WN
authors.lastname: Konings
authors.forename: J
authors.initials: J
authors.lastname: Kok
authors.forename: G
authors.initials: G
authors.lastname: Venema
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D015153:Blotting, Western; D004269:DNA, Bacterial; D013294:Lactococcus lactis / Q000201:enzymology* / Q000235:genetics; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011499:Protein Processing, Post-Translational; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008074:Lipoproteins; D012697:Serine Endopeptidases