title: Cleavage of Treponema denticola PrcA polypeptide to yield protease complex-associated proteins Prca1 and Prca2 is dependent on PrtP.
abstract: Analysis of potential virulence factors of oral spirochetes focuses on surface and secreted proteins. The Treponema denticola chymotrypsin-like protease (CTLP) is implicated in degradation of host cell molecules and contributes to tissue invasion. The CTLP complex, composed of the 72-kDa PrtP protein and two auxiliary proteins with molecular masses of approximately 40 and 30 kDa, is also involved in localization and oligomerization of the T. denticola major surface protein (Msp). The larger auxiliary protein was reported to be encoded by an open reading frame (ORF2) directly upstream of prtP. The deduced 39-kDa translation product of ORF2 contains a sequence matching the N-terminal sequence determined from one of the CTLP complex proteins. No proteins with significant homology are known, nor was information available on the third protein of the complex. DNA sequence analysis showed that ORF2 extended an additional 852 bp upstream of the reported sequence. The complete gene, designated prcA, encodes a predicted N-terminally-acylated polypeptide of approximately 70 kDa. Isogenic mutants with mutations in prtP, prcA, and prcA-prtP all lacked CTLP protease activity. The prcA mutant lacked all three CTLP proteins. The prcA-prtP mutant produced only a C-terminally-truncated 62-kDa PrcA protein. The prtP mutant produced a full-length 70-kDa PrcA. Immunoblot analysis of recombinant PrcA constructs confirmed that PrcA is cleaved to yield the two smaller proteins of the CTLP complex, designated PrcA1 and PrcA2. These data indicate that PrtP is required for cleavage of PrcA and suggest that this cleavage may be required for formation or stability of outer membrane complexes.
authors.affiliation: Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor 48109-1078, USA.
authors.forename: Si Young
authors.initials: SY
authors.lastname: Lee
authors.forename: Xue-Lin
authors.initials: XL
authors.lastname: Bian
authors.forename: Grace W K
authors.initials: GW
authors.lastname: Wong
authors.forename: Pauline M
authors.initials: PM
authors.lastname: Hannam
authors.forename: Barry C
authors.initials: BC
authors.lastname: McBride
authors.forename: J Christopher
authors.initials: JC
authors.lastname: Fenno
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D002918:Chymotrypsin / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases; D013381:Subtilisins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014208:Treponema / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C102527:PrcA protein, bacteria; D010447:Peptide Hydrolases; D013381:Subtilisins; C102570:dentilisin; D002918:Chymotrypsin