Enzymatic properties of dipeptidyl aminopeptidase IV produced by the periodontal pathogen Porphyromonas gingivalis and its participation in virulence.
Porphyromonas gingivalis is a major pathogen associated with adult periodontitis. We cloned and sequenced the gene (dpp) coding for dipeptidyl aminopeptidase IV (DPPIV) from P. gingivalis W83, based on the amino acid sequences of peptide fragments derived from purified DPPIV. An Escherichia coli strain overproducing P. gingivalis DPPIV was constructed. The enzymatic properties of recombinant DPPIV purified from the overproducer were similar to those of DPPIV isolated from P. gingivalis. The three amino acid residues Ser, Asp, and His, which are thought to form a catalytic triad in the C-terminal catalytic domain of eukaryotic DPPIV, are conserved in P. gingivalis DPPIV. When each of the corresponding residues of the enzyme was substituted with Ala by site-directed mutagenesis, DPPIV activity significantly decreased, suggesting that these three residues of P. gingivalis DPPIV are involved in the catalytic reaction. DPPIV-deficient mutants of P. gingivalis were constructed and subjected to animal experiments. Mice injected with the wild-type strain developed abscesses to a greater extent and died more frequently than those challenged with mutant strains. Mice injected with the mutants exhibited faster recovery from the infection, as assessed by weight gain and the rate of lesion healing. This decreased virulence of mutants compared with the parent strain suggests that DPPIV is a potential virulence factor of P. gingivalis and may play important roles in the pathogenesis of adult periodontitis induced by the organism.
Kumagai
Y
Y
Department of Microbiology, Nippon Dental University, Fujimi 1-9-20, Chiyoda-ku, Tokyo 102-8159, Japan. yumi-mic@tokyo.ndu.ac.jp
Konishi
K
K
Gomi
T
T
Yagishita
H
H
Yajima
A
A
Yoshikawa
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D018819:Dipeptidyl Peptidase 4 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D016966:Porphyromonas gingivalis / Q000201:enzymology* / Q000472:pathogenicity; D011994:Recombinant Proteins / Q000378:metabolism; D014774:Virulence
D011994:Recombinant Proteins; D018819:Dipeptidyl Peptidase 4