Activation of human prothrombin by arginine-specific cysteine proteinases (Gingipains R) from porphyromonas gingivalis.

Abstract

The effect of 95- (HRgpA) and 50-kDa gingipain R (RgpB), arginine-specific cysteine proteinases from periodontopathogenic bacterium Porphyromonas gingivalis on human prothrombin activation was investigated. Each enzyme released thrombin from prothrombin in a dose- and time-dependent manner with the former enzyme, containing adhesion domains, being 17-fold more efficient than the single chain RgpB. A close correlation between the generation of fibrinogen clotting activity and amidolytic activity indicated that alpha-thrombin was produced by gingipains R, and this was confirmed by SDS-polyacrylamide gel electrophoresis, thrombin active site labeling, and amino-terminal sequence analysis of prothrombin digestion fragments. Significantly, the catalytic efficiency of HRgpA to generate thrombin (k(cat)/K(m) = 1.2 x 10(6) m(-)1 s(-)1) was 100-fold higher than that of RgpB (k(cat)/K(m) = 1.2 x 10(4) m(-)1 s(-)1). The superior prothrombinase activity of HRgpA over RgpB correlates with the fact that only the former enzyme was able to clot plasma, and kinetic data indicate that prothrombin activation can occur in vivo. At P. gingivalis-infected periodontitis sites HRgpA may be involved in the direct production of thrombin and, therefore, in the generation of prostaglandins and interleukin-1, both have been found to be associated with the development and progression of the disease. Furthermore, by taking into account that the P. gingivalis bacterium has been immunolocalized in carotid atherosclerotic plaques at thrombus formation sites (Chiu, B. (1999) Am. Heart J. 138, S534-S536), our results indicate that bacterial proteinases may potentially participate in the pathogenesis of cardiovascular disease associated with periodontitis.

• Authors: T Imamura, A Banbula, P J Pereira, J Travis, J Potempa
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 276 Issue 22 Pg. 18984-91 (Jun 01 2001) ISSN: 0021-9258 [Print] United States
• PMID: 11278315 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Adhesins, Bacterial
• Gingipain Cysteine Endopeptidases
• Hemagglutinins
• Interleukin-1
• Prothrombin
• Fibrinogen
• Arginine
• Endopeptidases
• Thrombin
• Cysteine Endopeptidases

Topics

• Adhesins, Bacterial
• Arginine (metabolism)
• Binding Sites
• Blood Coagulation
• Blotting, Western
• Cell Adhesion
• Cysteine Endopeptidases (chemistry, metabolism)
• Dose-Response Relationship, Drug
• Electrophoresis, Polyacrylamide Gel
• Endopeptidases (metabolism)
• Enzyme Activation
• Fibrinogen (metabolism)
• Gingipain Cysteine Endopeptidases
• Hemagglutinins (chemistry)
• Humans
• Interleukin-1 (biosynthesis)
• Kinetics
• Plasma (metabolism)
• Porphyromonas gingivalis (metabolism)
• Prothrombin (chemistry, metabolism)
• Sequence Analysis, Protein
• Thrombin (chemistry, metabolism)
• Time Factors