Differential cleavage of surface proteins of Borrelia burgdorferi by proteases.
The differential cleavage of surface proteins of Borrelia burgdorferi IRS strains by several proteases was examined. Proteinase K, trypsin, chymotrypsin and thermolysin all cleaved the outer surface protein B (OspB) to undetectable levels by Coomassie Brilliant Blue staining, whereas some residual protein was detected by immunoblotting with polyclonal and monoclonal antibodies. Not even antigenic fragments were detectable by immunoblotting with 1A8 monoclonal antibody reactive with OspB. Less effective or ineffective was the cleavage of OspB by V8 protease and proteinase A, respectively. The outer surface protein A was cleaved only by proteinase K. The effect of trypsin on borreliae viability and adhesion to cultured cells was also studied. The trypsin treatment of borreliae did not impair the viability of organisms which continued to synthesize the cleaved OspB. The attachment of B. burgdorferi to HEp-2 cells was reduced by 41% after treatment with trypsin, whereas preincubation of borreliae with monoclonal antibody 1A8 and guinea pig immune serum reduced the adhesion of borreliae to the cells by 32% and 87%, respectively.
Moroni
A
A
Istituto di Microbiologia, University of Bologna, Italy.
Sambri
V
V
Massaria
F
F
La Placa
M
M
Brocchi
E
E
De Simone
F
F
Cevenini
R
R
D000911:Antibodies, Monoclonal; D001287:Attachment Sites, Microbiological; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D015748:Borrelia burgdorferi Group / Q000737:chemistry* / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D002918:Chymotrypsin / Q000378:metabolism; D019286:Endopeptidase K; D010450:Endopeptidases / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D012697:Serine Endopeptidases / Q000378:metabolism; D013820:Thermolysin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D010450:Endopeptidases; D012697:Serine Endopeptidases; D002918:Chymotrypsin; D019286:Endopeptidase K; D013820:Thermolysin