Purification and characterization of a 50-kDa cysteine proteinase (gingipain) from Porphyromonas gingivalis.
Porphyromonas gingivalis, a Gram-negative anaerobic rod, has been closely associated with the initiation and progression of periodontal disease. This organism has been shown to produce a large number of proteolytic enzymes which can degrade a variety of tissue proteins, and these are considered to be major virulence factors. One of the proteinases produced by this organism, referred to as gingipain-1, has been purified to homogeneity from P. gingivalis culture medium by a combination of gel filtration and ion-exchange chromatography. The enzyme was found to have a molecular mass near 50 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a pH optimum in the neutral to alkaline range, and a requirement for cysteine for activation and Ca2+ for stabilization. Amino-terminal sequence analysis indicated that gingipain belongs to a new, so far unknown, subfamily of cysteine proteinases. Three unusual features of this proteinase are: (a) the stimulation of amidolytic activity by glycine-containing dipeptides; (b) a narrow specificity which is limited to peptide bonds containing arginine residues; and (c) resistance to inhibition by proteinase inhibitors in human plasma.
Chen
Z
Z
Department of Biochemistry, University of Georgia, Athens 30602.
Potempa
J
J
Polanowski
A
A
Wikstrom
M
M
Travis
J
J
D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D001439:Bacteroides / Q000201:enzymology*; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data
D018829:Adhesins, Bacterial; D002413:Cations, Divalent; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D003546:Cysteine Endopeptidases; D002118:Calcium