title: Lysine- and arginine-specific proteinases from Porphyromonas gingivalis. Isolation, characterization, and evidence for the existence of complexes with hemagglutinins.
abstract: Porphyromonas gingivalis contains many virulence factors that have been implicated as participants in the progression of periodontal disease. It has been shown to produce proteinases of "trypsin-like" specificity in a number of molecular forms, but previous work in our laboratory resulted in the purification of a major arginine-specific cysteine proteinase, gingipain, which contradicted this supposed specificity. In this study, separate proteinases with arginine and lysine specificity were isolated from a high molecular mass fraction of the P. gingivalis culture fluid. The arginine-specific enzyme was found, by amino acid sequencing studies, to be a high molecular mass form of gingipain, formed by the 50-kDa gingipain noncovalently complexed with 44-kDa binding proteins, subsequently identified as hemagglutinins. The 60-kDa lysine-specific proteinase, referred to as Lys-gingipain, was also found to have one of these hemagglutinins complexed with it in the same manner. Lys-gingipain was found to be a cysteine proteinase with optimal activity and stability at pH 8.0-8.5 and was extensively characterized in terms of its specificity and activation characteristics. The proteinase-hemagglutinin complexes may be important in the uptake of hemin, a vital metabolite for P. gingivalis, via hemagglutination and subsequent hemolysis of erythrocytes.
authors.affiliation: Department of Biochemistry, University of Georgia, Athens 30602.
authors.forename: R
authors.initials: R
authors.lastname: Pike
authors.forename: W
authors.initials: W
authors.lastname: McGraw
authors.forename: J
authors.initials: J
authors.lastname: Potempa
authors.forename: J
authors.initials: J
authors.lastname: Travis
mesh_terms: D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D001120:Arginine / Q000378:metabolism*; D002850:Chromatography, Gel; D003546:Cysteine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism*; D008969:Molecular Sequence Data; D016966:Porphyromonas gingivalis / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity
chemical_list: D018829:Adhesins, Bacterial; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D011480:Protease Inhibitors; D001120:Arginine; D003546:Cysteine Endopeptidases; D008239:Lysine