Structural basis for reduced staphylocoagulase-mediated bovine prothrombin activation

Rainer Friedrich, Peter Panizzi, Shun Ichiro Kawabata, Wolfram Bode, Paul E. Bock, Pablo Fuentes-Prior

研究成果: ジャーナルへの寄稿学術誌査読

17 被引用数 (Scopus)

抄録

Staphylocoagulase (SC) is a protein secreted by the human pathogen, Staphylococcus aureus, that activates human prothrombin (ProT) by inducing a conformational change. SC-bound ProT efficiently clots fibrinogen, thus bypassing the physiological blood coagulation pathway. The crystal structure of a fully active SC fragment, SC-(1-325), bound to human prethrombin 2 showed that the SC-(1-325) N terminus inserts into the Ile16 pocket of prethrombin 2, thereby inducing expression of a functional catalytic site in the cognate zymogen without peptide bond cleavage. As shown here, SC-(1-325) binds to bovine and human ProT with similar affinity but activates the bovine zymogen only very poorly. By contrast to the ∼2-fold difference in chromogenic substrate kinetic constants between human thrombin and the SC-(1-325) ·human (pro)thrombin complexes, SC-(1-325)·bovine ProT shows a 3,500-fold lower kcat/Km compared with free bovine thrombin, because of a 47-fold increase in Km and a 67-fold decrease in kcat. The SC-(1-325)·bovine ProT complex is ∼5,800-fold less active compared with its human counterpart. Comparison of human and bovine fibrinogen as substrates of human and bovine thrombin and the SC-(1-325)·(pro)thrombin complexes indicates that the species specificity of SC-(1-325) cofactor activity is determined primarily by differences in conformational activation of bound ProT. These results suggest that the catalytic site in the SC-(1-325)·bovine ProT complex is incompletely formed. The current crystal structure of SC-(1-325)·bovine thrombin reveals that SC would dock similarly to the bovine proenzyme, whereas the bovine (pro)thrombin-characteristic residues Arg144 and Arg145 would likely interfere with insertion of the SC N terminus, thus explaining the greatly reduced activation of bovine ProT.

本文言語英語
ページ(範囲)1188-1195
ページ数8
ジャーナルJournal of Biological Chemistry
281
2
DOI
出版ステータス出版済み - 1月 13 2006

!!!All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学
  • 細胞生物学

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