Nucleotide and deduced protein sequence of the extracellular, serine basic protease gene (bprB) from Dichelobacter nodosus strain 305: comparison with the basic protease gene (bprV) from virulent strain 198.
In earlier studies, it appeared that benign strains of the Gram-negative, obligate anaerobe, Dichelobacter nodosus, were devoid of the extracellular, serine basic protease (pI approximately 9.5) of virulent strains. However, Southern and PCR analysis have shown a homologous gene (bprB) in the representative benign strain 305. The deduced amino acid sequence of the prepro- and mature protease regions of bprB confirmed this homology and showed 97% sequence identity with the bprV precursor from virulent strain 198. Identity in the carboxy-terminal extension region was 90%. Expression studies in Escherichia coli transformed with bprB, showed that the gene was capable of the production of an active protease. A protease, albeit with a lower iso-electric point (approximately 8.6), was isolated from D. nodosus culture supernatants and shown to cross-react with antibodies raised against the more basic protease from strain 198. The amino acid sequence encoded by the strain 305 gene revealed two additional acidic residues consistent with a lowered iso-electric point and supported the conclusion that bprB and bprV produce equivalent basic proteases.
Lilley
G G
GG
CSIRO, Division of Biomolecular Engineering, Parkville, Victoria, Australia.
Riffkin
M C
MC
Stewart
D J
DJ
Kortt
A A
AA
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D017931:DNA Primers; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics*; D014774:Virulence / Q000235:genetics
D001426:Bacterial Proteins; D017931:DNA Primers; D012697:Serine Endopeptidases; C078345:extracellular basic protease, Dichelobacter nodosus