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Characterization of N‐CBz‐glycyl‐glycyl‐arginyl peptidase and glycyl‐prolyl peptidase of Bacteroides gingivalis
Author(s) -
Suido H.,
Neiders M. E.,
Barua P. K.,
Nakamura M.,
Mashimo P. A.,
Genco R. J.
Publication year - 1987
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/j.1600-0765.1987.tb01608.x
Subject(s) - leupeptin , antipain , enzyme , biochemistry , bacteroides , protease , chemistry , proteases , serine protease , bacteroidaceae , tmprss6 , microbiology and biotechnology , biology , bacteria , genetics
Bacteroides gingivalis produces large amounts of proteolytic enzymes which may play a role in its virulence. These enzymes may participate in the tissue destruction of the inflammatory process. In this study, the characteristics of two such enzymes, N‐CBz‐glycyl‐glycyl‐arginyl peptidase (N‐CBz‐Gly‐Gly‐Arg peptidase) and glycyl‐prolyl peptidase (Gly‐Pro peptidase) were investigated. The enzymes eluted in different peaks from an anion exchange column. N‐CBz‐Gly‐Gly‐Arg peptidase was associated with cells up to 48 h in culture. If cultured longer, it also released in the supernatant. It exhibited optimal activity between pH of 7.0 and 7.5 and was readily inactivated by heat treatment (45°C for 15 min). The enzyme activity was inhibited by p‐chloromercuribenzoic acid (PCMB), leupeptin and antipain, suggesting that it is a thiol protease. The B. gingivalis N‐CBz‐Gly‐Gly‐Arg peptidase was different from the serum enzyme that digests the same substrate. The serum enzyme was more resistant to heat treatment and was inhibited by diisopro‐pylfluorophosphate (DFP). B. gingivalis also produced Gly‐Pro peptidase that is released in the supernatant. The enzyme has an optimal pH range between 7.5 and 8.0. The B. gingivalis Gly‐Pro peptidase was inhibited by DFP, suggesting that it represents a serine protease. The serum Gly‐Pro peptidase did not differ from the bacterial enzyme with respect to its sensitivity to inhibitors; however, they were markedly different in heat sensitivity. The bacterial enzyme was completely inactivated at 60°C for 30 min, whereas the serum enzyme was not inactivated even at 1 h at 60°C.