The opdB locus encodes the trypsin-like peptidase activity of Treponema denticola.
High levels of Treponema denticola in subgingival dental plaque are associated with severe periodontal disease. T. denticola, along with Porphyromonas gingivalis and Bacteroides forsythus, are the only cultivatable oral microorganisms that produce significant amounts of "trypsin-like" peptidase activity. The ability of subgingival plaque to hydrolyze N-alpha-benzoyl-DL-arginine-2-naphthylamide (BANA) is associated with high levels of one or more of these organisms. The purpose of this study was to identify the gene encoding trypsin-like activity in T. denticola and thus facilitate molecular-level studies of its potential role in disease. Using published peptide sequences of a T. denticola surface-associated oligopeptidase with BANA-hydrolyzing activity, we identified the gene, designated opdB, in an apparently noncoding region of the T. denticola genome unannotated contigs (11/2000; http://www.tigr.org). The opdB gene begins with a TTG start codon and encodes a 685-residue peptide with high homology to the oligopeptidase B family in prokaryotes and eukaryotes. An isogenic T. denticola opdB mutant was constructed by allelic replacement mutagenesis using an ermF/AM gene cassette. The mutant lacked BANA-hydrolyzing activity and had a slightly slower growth rate than the parent strain. This mutant will be used in future studies of interactions of T. denticola with host cells and tissue.
Fenno
J C
JC
Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, Michigan 48109-1078, USA. fenno@umich.edu
Lee
S Y
SY
Bayer
C H
CH
Ning
Y
Y
D000483:Alleles; D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D001587:Benzoylarginine-2-Naphthylamide / Q000378:metabolism; D004269:DNA, Bacterial; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D014208:Treponema / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D001587:Benzoylarginine-2-Naphthylamide; D012697:Serine Endopeptidases; C090703:oligopeptidase B