Cleavage of human complement component C5 by cysteine proteinases from Porphyromonas (Bacteroides) gingivalis. Prior oxidation of C5 augments proteinase digestion of C5.
Since severe periodontitis is characterized by an acute inflammatory response with cellular infiltration and microbial overgrowth, plasma proteins could be exposed to both proteinases and oxidants released from the granulocytes, as well as to proteinases from the microorganisms. When human complement component C5 was digested by cysteine proteinases (i.e. gingipain-R and gingipain-K) from Porphyromonas gingivalis, limited cleavage of the C5 molecule was observed. If C5 was first oxidized by hydroxyl radicals, these gingipains converted modified C5 to fragments that exhibited significantly greater pro-inflammatory activity than did digests of unmodified C5. After cleavage of oxidized C5 by gingipain-R, the digest exhibited measurably greater neutrophil enzyme release and chemotaxis of human polymorphonuclear leukocytes (PMNs) compared with the activities of unoxidized C5 digests. Gingipain-K generates virtually no polarization or chemotactic activity of human PMNs from C5, nor is enzyme release stimulated by these C5 digests. However, when oxidized C5 was digested by gingipain-K, human PMNs were stimulated for polarization, chemotaxis and enzyme release indicating that an active fragment had been generated. Proteolysis of oxidized C5 evokes greater neutrophil activation than does proteolysis of unoxidized protein, a fact which supports the hypothesis that oxidation and proteolysis may be coupled to enhance the destructive effects of the inflammatory process. These results, in which digests of both oxidized and unmodified complement component C5 were evaluated, support the general concept that oxidation and proteolysis may participate cooperatively in amplifying both the severity and duration of the inflammatory reaction.
Discipio
R G
RG
La Jolla Institute for Experimental Medicine, CA, USA.
Daffern
P J
PJ
Kawahara
M
M
Pike
R
R
Travis
J
J
Hugli
T E
TE
Potempa
J
J
D018829:Adhesins, Bacterial; D018929:Cell Culture Techniques; D002634:Chemotaxis, Leukocyte; D003176:Complement C3 / Q000378:metabolism; D003182:Complement C5 / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins / Q000276:immunology*; D006801:Humans; D007700:Kinetics; D018375:Neutrophil Activation / Q000276:immunology*; D010084:Oxidation-Reduction; D016966:Porphyromonas gingivalis / Q000201:enzymology*
D018829:Adhesins, Bacterial; D003176:Complement C3; D003182:Complement C5; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D003546:Cysteine Endopeptidases