Proteolytic activity among various oral Treponema species and cloning of a prtP-like gene of Treponema socranskii subsp. socranskii.
The proteolytic activity of 11 treponemal strains representing different phylogenetic groups was investigated by SDS-polyacrylamide gel electrophoresis with copolymerised casein, gelatin and fibrinogen as substrates. The activity was specified to be trypsin- and chymotrypsin-like by the cleavage of synthetic substrates BAPNA and SAAPFNA, respectively. Nine strains degrade casein and the synthetic substrate BAPNA. Chymotrypsin-like activity specifically inhibited by phenylmethylsulfonyl fluoride was found in four treponemes. Southern blot analysis using a Treponema socranskii subsp. socranskii-specific prtP probe confirmed the presence of prtP homologous genes in these four strains. The internal fragments of the chymotrypsin-like protease genes were cloned and sequenced after PCR amplification. Here we report the cloning of the complete prtP-like gene of T. socranskii subsp. socranskii, an organism shown to possess epidemiologic relevance in periodontitis.
Heuner
K
K
Institut für Moleculare Infektionsbiologie der Universität Würzburg, Germany. klaus.heuner@mail.uni-wuerzburg.de
Bergmann
I
I
Heckenbach
K
K
Göbel
U B
UB
D015139:Blotting, Southern; D002918:Chymotrypsin / Q000737:chemistry; D003001:Cloning, Molecular; D018093:Gelatinases / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial / Q000235:genetics; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010518:Periodontitis / Q000382:microbiology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D013379:Substrate Specificity; D014208:Treponema / Q000145:classification / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D014357:Trypsin / Q000737:chemistry
D012697:Serine Endopeptidases; D002918:Chymotrypsin; D014357:Trypsin; C085783:lactocepin; D018093:Gelatinases