Journal: Journal of Bacteriology
Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex ▿
Figure Lengend Snippet: Interaction of PrcB with PrtP. Arrows denote PrcB, PrtP, and rabbit IgG heavy chains (H). (A) Immunoblots probed with anti-PrtP antibodies and HisProbe reagent, as indicated. Lanes 1 and 2, T. denticola CF499; lanes 3 and 4, CF417; lanes 1 and 4, cell extracts; lanes 2 and 3, anti-PrtP immunoprecipitates from cell extracts. (B) T. denticola CF499 lysates were immunoprecipitated with polyclonal antibodies to PrtP, PrcA, FlaA, or Msp. Eluted proteins on duplicate blots were probed with anti-PrtP antibodies and HisProbe reagent, as indicated. (C) Immunoblot probed with anti-PrtP antibodies. Lanes 1 to 3, T. denticola CF499; lanes 4 to 6, T. denticola CF417; lanes 1 and 4, whole-cell extracts; lanes 2 and 5, anti-6×His monoclonal antibody immunoprecipitates from cell extracts; lanes 3 and 6, anti-PrtP antibody immunoprecipitates from cell extracts. (D) E. coli cells expressing PrtP, 6×His-PrcB, or both PrtP and 6×His-PrcB were analyzed directly and after Ni 2+ chromatography. Immunoblots of whole-cell extracts and Ni 2+ affinity column eluates were probed with anti-6×His monoclonal antibody or anti-PrtP polyclonal antibodies. Lanes: 1, E. coli /pCF411 (expresses PrtP); 2, E. coli /pCF415 (expresses PrcB-6×His); 3, E. coli /pCF416 (expresses PrcB-6×His + PrtP).
Article Snippet: Annotation of the prcB open reading frame was done using the FGENESB algorithm trained to the T. denticola genome sequence (Softberry, Inc., Mt.
Techniques: Western Blot, Immunoprecipitation, Expressing, Chromatography, Affinity Column