Hydrolysis of the Leu-Gly bond of phenylazobenzyl-oxycarbonyl- l -Pro- l -Leu-Gly- l -Pro- D -Arg (a substrate of microbial collagenases) by treponemes isolated from the subgingival plaque of periodontitis patients

Abstract

Cell extracts prepared from several oral treponemes isolated from the subgingival plaque of periodontitis patients showed high enzyme activity toward phenylazobenzyl-oxycarbonyl- l -prolyl- l -leucylglycyl- l -prolyl- d -arginine (a compound used as a substrate for microbial collagenases). One major enzyme hydrolyzing this substrate at the Leu-Gly bond only was partially purified from an unspeciated treponeme (strain US), Treponema denticola ATCC 35405, and 29 different clinical isolates of T. denticola . The Treponema US enzyme also hydrolyzed furylacryloyl- l -leucylglycyl- l -prolyl- l -alanine (another substrate of bacterial collagenases) at the Leu-Gly bond. This enzyme also hydrolyzed various collagens and collagen-derived peptides. These treponemal proteases were sensitive to metal chelators and p -chloromercury compounds. The results indicate that human oral treponemes contain enzymes that readily hydrolyze in chromogenic protease substrates the Leu-Gly bond only that is the cleavage site of these substrates also by “true” microbial collagenases.