title: Treponema denticola PrcB is required for expression and activity of the PrcA-PrtP (dentilisin) complex.
abstract: The Treponema denticola surface protease complex, consisting of PrtP protease (dentilisin) and two auxiliary polypeptides (PrcA1 and PrcA2), is believed to contribute to periodontal disease by degrading extracellular matrix components and disrupting host intercellular signaling. Previously, we showed that transcription of the protease operon initiates upstream of TDE0760 (herein designated prcB), the open reading frame immediately 5' of prcA-prtP. The prcB gene is conserved in T. denticola strains. PrcB localizes to the detergent phase of Triton X-114 cell surface extracts and migrates as a 22-kDa polypeptide, in contrast to the predicted 17-kDa cytoplasmic protein encoded in the annotated T. denticola genome. Consistent with this observation, the PrcB N terminus is unavailable for Edman sequencing, suggesting that it is acylated. Nonpolar deletion of prcB in T. denticola showed that PrcB is required for production of PrtP protease activity, including native PrtP cleavage of PrcA to PrcA1 and PrcA2. A 6xHis-tagged PrcB protein coimmunoprecipitates with native PrtP, using either anti-PrtP or anti-His-tag antibodies, and recombinant PrtP copurifies with PrcB-6xHis in nickel affinity chromatography. Taken together, these data are consistent with identification of PrcB as a PrtP-binding lipoprotein that likely stabilizes the PrtP polypeptide during localization to the outer membrane.
authors.affiliation: Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, MI 48109-1078, USA.
authors.forename: Valentina
authors.initials: V
authors.lastname: Godovikova
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Wang
authors.forename: M Paula
authors.initials: MP
authors.lastname: Goetting-Minesky
authors.forename: Yu
authors.initials: Y
authors.lastname: Ning
authors.forename: Ricardo F
authors.initials: RF
authors.lastname: Capone
authors.forename: Claudia K
authors.initials: CK
authors.lastname: Slater
authors.forename: J Christopher
authors.initials: JC
authors.lastname: Fenno
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D002918:Chymotrypsin / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases; D013381:Subtilisins / Q000235:genetics / Q000378:metabolism*; D048373:Treponema denticola / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C102527:PrcA protein, bacteria; D010447:Peptide Hydrolases; D013381:Subtilisins; C102570:dentilisin; D002918:Chymotrypsin