Cytopathic effects of the major surface protein and the chymotrypsinlike protease of Treponema denticola.
Prominent antigens of Treponema denticola have been suggested to be mediators of the cytopathic effects typically seen in periodontal disease. In the present study of the T. denticola major surface protein (Msp) and the surface-expressed chymotrypsinlike protease complex (CTLP), we characterized the ability of these proteins to adhere to and lyse epithelial cells. Msp and CTLP were closely associated in spirochete outer membranes. Purified Msp, both native and recombinant, and CTLP bound to glutaraldehyde-fixed periodontal ligament epithelial cells. Adherence of Msp was partially blocked by specific antibodies. Adherence of CTLP was partially blocked by serine protease inhibitors and was further inhibited by specific antibodies. Both native Msp and CTLP were cytotoxic toward periodontal ligament epithelial cells, and their cytotoxicity was inhibited by the same treatments that inhibited adherence. Msp, but not CTLP, lysed erythrocytes. Msp complex (partially purified outer membranes free of protease activity) was cytotoxic toward a variety of different cell types. Pore-forming activities of recombinant Msp in black lipid model membrane assays and in HeLa cell membranes were similar to those reported for the native protein, supporting the hypothesis that Msp cytotoxicity was due to its pore-forming activity.
Fenno
J C
JC
Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada.
Hannam
P M
PM
Leung
W K
WK
Tamura
M
M
Uitto
V J
VJ
McBride
B C
BC
D000818:Animals; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification / Q000633:toxicity*; D016466:CHO Cells; D053818:Chymases; D006224:Cricetinae; D006367:HeLa Cells; D006461:Hemolysis; D006801:Humans; D008970:Molecular Weight; D018272:Porins / Q000633:toxicity; D011817:Rabbits; D012697:Serine Endopeptidases / Q000633:toxicity*; D013552:Swine; D014208:Treponema / Q000472:pathogenicity*
D001425:Bacterial Outer Membrane Proteins; D018272:Porins; D012697:Serine Endopeptidases; D053818:Chymases