Activation of blood coagulation factor IX by gingipains R, arginine-specific cysteine proteinases from Porphyromonas gingivalis

Imamura, Takahisa; Tanase, Sumio; Hamamoto, Takayoshi; Potempa, Jan; Travis, James

doi:10.1042/0264-6021:3530325

Cited by 26 publications

(25 citation statements)

References 50 publications

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“…As a matter of fact, studies have shown that P. gingivalis Arg-gingipains are able to activate prothrombin and coagulation factor X. 39,40 Interestingly, PAR 2 has been described as the endogenous receptor mediating the effects of factor Xa in endothelial cells. 41 Therefore, P. gingivalis Arg-gingipains could account for PAR 2 activation both directly (by cleaving the receptor) and indirectly (by inducing the release of PAR 2 -activating proteases).…”

Section: Discussionmentioning

confidence: 99%

Protease-Activated Receptor-2 Activation

Holzhausen

Spolidorio

Ellen

et al. 2006

The American Journal of Pathology

101

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We have investigated the specific contribution of protease-activated receptor-2 (PAR 2 ) to host defense during Porphyromonas gingivalis infection. Culture supernatants from P. gingivalis strains 33277 and W50 provoked Ca 2؉ mobilization in cells transfected with PAR 2 (PAR 2 -KNRK) and desensitized the subsequent responses to PAR 2 -selective agonist. In addition, culture supernatants of P. gingivalis E8 (RgpA/RgpB double knockout) did not cause calcium response in PAR 2 -KNRK cells, evidencing the involvement of the arginine-specific cysteine proteases RgpA and RgpB in PAR 2 activation by P. gingivalis. Injection of P. gingivalis into mouse subcutaneous chambers provoked an increased proteolytic activity, which was inhibited by serine protease inhibitors. Fluids collected from chambers of P. gingivalis-injected mice were able to activate PAR 2 and this activation was inhibited by serine protease inhibitors. P. gingivalis inoculation into subcutaneous chambers of wild-type mice induced an inflammatory response that was inhibited by a serine protease inhibitor and was significantly reduced in PAR 2 -deficient mice. Finally, mice orally challenged with P. gingivalis developed alveolar bone loss, which was significantly reduced in PAR 2 -deficient mice at 42 and 60 days after P. gingivalis infection. We conclude that PAR 2 is activated on P. gingivalis infection, in which it plays an important role in the host inflammatory response. (Am J Pathol

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Section: Discussionmentioning

confidence: 99%

Protease-Activated Receptor-2 Activation

Holzhausen

Spolidorio

Ellen

et al. 2006

The American Journal of Pathology

101

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“…In comparison to rgpA, the rgpB gene lacks a sequence encoding the hemagglutinin/ adhesin domains, and its product occurs predominantly as the soluble 50 kDa gingipain-R (RgpB) (13). Gingipain-R enzymes have been shown to activate coagulation factors and degrade components of the complement pathway and several physiologically important proteins, contributing to the virulence of the pathogen (14)(15)(16)(17)(18)(19). In addition to causing destruction to the tissue supporting the teeth, gingipain-R enzymes play a major role in the deregulation of the inflammatory response and disruption of host defense mechanisms.…”

mentioning

confidence: 99%

Characterization of the Specificity of Arginine-Specific Gingipains from Porphyromonas gingivalis Reveals Active Site Differences between Different Forms of the Enzymes

et al. 2003

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Porphyromonas gingiValis is a pathogen associated with periodontal disease, and argininespecific proteases (gingipains-R) from the bacterium are important virulence factors. The specificity of two forms of gingipain-R, HRgpA and RgpB, for substrate positions C-terminal to the cleavage site was analyzed, and notable differences were observed between the enzymes. Molecular modeling of the HRgpA catalytic domain, based on the structure of RgpB, revealed that there are four amino acid substitutions around the active site of HRgpA relative to RgpB that may explain their different specificity. Previously, differences in the ability of these two gingipain-R forms to cleave a number of proteins were attributed to additional adhesins on HRgpA mediating increased interaction with the substrates. Here, purified RgpA cat , the catalytic domain of HRgpA, which like RgpB also lacks adhesin subunits, was used to show that the differences between HRgpA and RgpB are probably due to the amino acid substitutions at the active site. The kinetics of cleavage of fibrinogen, a typical protein substrate for the gingipain-R enzymes, which is bound by HRgpA but not RgpA cat or RgpB, were evaluated, and it was shown that there was no difference in the cleavage of the fibrinogen AR-chain between the different enzyme forms. HRgpA degraded the fibrinogen B -chain more efficiently, generating distinct cleavage products. This indicates that while the adhesin domain(s) play(s) a minor role in the cleavage of protein substrates, the major effect is still provided by the amino acid substitutions at the active site of rgpA gene products versus those of the rgpB gene.

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“…In addition to the ability to generate directly ␣-thrombin from prothrombin, gingipains R are capable of efficiently activating factors X and IX and accelerating in this way plasma coagulation through the cascade reaction of the clotting factors in the presence of phospholipids and Ca 2ϩ (32,40). In order to understand the relative importance of these three activities, we have directly compared the ability of HRgpA to promote the clotting of normal plasma versus factor X-, IX-, and XI-deficient plasmas by measuring APTT.…”

Section: Activation Of Human Prothrombin By Gingipains R-in Or-mentioning

confidence: 99%

Activation of Human Prothrombin by Arginine-specific Cysteine Proteinases (Gingipains R) from Porphyromonas gingivalis *

Imamura

Bańbuła

Pereira

et al. 2001

Journal of Biological Chemistry

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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 ␣-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 ؋ 10 6 M ؊1 s ؊1 ) was 100-fold higher than that of RgpB (k cat /K m ‫؍‬ 1.2 ؋ 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.

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Activation of blood coagulation factor IX by gingipains R, arginine-specific cysteine proteinases from Porphyromonas gingivalis

Cited by 26 publications

References 50 publications

Protease-Activated Receptor-2 Activation

Protease-Activated Receptor-2 Activation

Characterization of the Specificity of Arginine-Specific Gingipains from Porphyromonas gingivalis Reveals Active Site Differences between Different Forms of the Enzymes

Activation of Human Prothrombin by Arginine-specific Cysteine Proteinases (Gingipains R) from Porphyromonas gingivalis *

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