論文

査読有り
2017年11月

In vitro exploration of latent prothrombin mutants conveying antithrombin resistance

THROMBOSIS RESEARCH
  • Shogo Tamura
  • ,
  • Moe Murata-Kawakami
  • ,
  • Yuki Takagi
  • ,
  • Sachiko Suzuki
  • ,
  • Akira Katsumi
  • ,
  • Akira Takagi
  • ,
  • Tetsuhito Kojima

159
開始ページ
33
終了ページ
38
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.thromres.2017.09.020
出版者・発行元
PERGAMON-ELSEVIER SCIENCE LTD

Introduction: Antithrombin resistance (ATR) prothrombinemia is an inherited thrombophilic disorder caused by missense mutations in prothrombin gene (F2) at Arg596 of the sodium-binding region. Previously, prothrombin mutants Yukuhashi (Arg596Leu), Belgrade (Arg596Gln), and Padua 2 (Arg596Trp) were reported as ATR-prothrombins possessing a risk of familial venous thrombosis. To identify additional F2 mutations causing the ATR-phenotype, we investigated the coagulant properties of recombinant prothrombins mutated at amino acid residues within the sodium-binding region by single nucleotide substitutions (Thr540, Arg541, Glu592, and Lys599).
Materials and methods: We constructed expression vectors of prothrombin mutants, established stably transfected HEK293 cells, and isolated the recombinant prothrombin proteins. We evaluated procoagulant activity and ATR-phenotypes of those mutants in reconstituted plasma by mixing with prothrombin deficient plasma.
Results: The secreted quantity of all prothrombin mutants was the same as that of the wild-type prothrombin. Procoagulant activity of each mutant varied from 1.7% to 79.5% in a one-stage clotting assay and from 2.0% to 104.5% in a two-stage chromogenic assay. Most prothrombin mutants tested presented with a severe ATR-phenotype. To estimate the thrombosis risk of these mutations, we determined the residual clotting activity (RCA) after 30 min inactivation with antithrombin. RCA scores, normalized to the wild-type, revealed that prothrombin mutants Lys599Arg (5.35) and Glu592Gln (4.71) had high scores, which were comparable with prothrombins Yukuhashi (4.36) and Belgrade (5.19).
Conclusions: Mutation of prothrombin at the sodium-binding site caused ATR-phenotypes. Of those tested, Lys599Arg and Glu592Gln may possess a thrombosis risk as large as the known pathogenic prothrombins Yukuhashi and Belgrade.

Web of Science ® 被引用回数 : 5

リンク情報
DOI
https://doi.org/10.1016/j.thromres.2017.09.020
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000418021100007&DestApp=WOS_CPL

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