Involvement of a lysine-specific cysteine proteinase in hemoglobin adsorption and heme accumulation by Porphyromonas gingivalis.
The oral anaerobic bacterium Porphyromonas gingivalis, a major pathogen of advanced adult periodontitis, produces a novel class of cysteine proteinases in both cell-associated and secretory forms. A lysine-specific cysteine proteinase (Lys-gingipain, KGP), as well as an arginine-specific cysteine proteinase (Arg-gingipain), is a major trypsin-like proteinase of the organism. Recent studies indicate that the secreted KGP is implicated in the destruction of periodontal tissue and the disruption of host defense mechanisms. In this study, we have constructed a KGP-deficient mutant to determine whether the cell-associated KGP is important for pathophysiology of the organism. Although the mutant retained the strong ability to disrupt the bactericidal activity of polymorphonuclear leukocytes, its hemagglutination activity was reduced to about one-half that observed with the wild-type strain. More important, the mutant did not form black-pigmented colonies on blood agar plates, indicating the defect of hemoglobin adsorption and heme accumulation. Immunoblot analysis showed that the expression of a 19-kDa hemoglobin receptor protein, which is thought to be responsible for hemoglobin binding by the organism, was greatly retarded in this mutant. The mutant also showed a marked decrease in the ability to degrade fibrinogen. These results suggest the possible involvement of KGP in the hemoglobin binding and heme accumulation of the organism and in the bleeding tendency in periodontal pockets.
Okamoto
K
K
Department of Pharmacology, Kyushu University Faculty of Dentistry, Higashi-ku, Fukuoka 812-8582, Japan.
Nakayama
K
K
Kadowaki
T
T
Abe
N
N
Ratnayake
D B
DB
Yamamoto
K
K
D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism; D000327:Adsorption; D001483:Base Sequence; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D015342:DNA Probes; D004269:DNA, Bacterial; D005340:Fibrinogen / Q000378:metabolism; D000080867:Gingipain Cysteine Endopeptidases; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000235:genetics / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism*; D006868:Hydrolysis; D008239:Lysine / Q000378:metabolism*; D009154:Mutation; D016966:Porphyromonas gingivalis / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000096:biosynthesis
D018829:Adhesins, Bacterial; D015342:DNA Probes; D004269:DNA, Bacterial; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D006454:Hemoglobins; D011956:Receptors, Cell Surface; D006418:Heme; D005340:Fibrinogen; D003546:Cysteine Endopeptidases; D008239:Lysine