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Detection of von Willebrand factor-cleaving protease (ADAMTS-13) in human platelets
Authors:Suzuki Misako  Murata Mitsuru  Matsubara Yumiko  Uchida Toshihiro  Ishihara Hiroaki  Shibano Toshiro  Ashida Shin-ichiro  Soejima Kenji  Okada Yasunori  Ikeda Yasuo
Affiliation:Department of Internal Medicine, Keio University, Tokyo 160-8582, Japan.
Abstract:The hemostatic activity of von Willebrand factor (vWF) is strongly dependent on its multimeric structure, with the highest activity in 'unusually large' multimers secreted from endothelial cells. The multimeric structure is regulated by a plasma protease, vWF-cleaving protease (vWF-CP, or ADAMTS-13). ADAMTS-13 mRNA is variably expressed in liver and other tissues. Because 15-25% of circulating vWF is stored in platelets, the presence and function of ADAMTS-13 in platelets are important issues. Here we report ADAMTS-13 expression in human platelets. Western blot analysis and flow cytometric analysis on permeabilized platelets revealed the presence of ADAMTS-13 protein. Real-time PCR demonstrated that ADAMTS-13 mRNA is present in platelets of six healthy volunteers, with little quantitative difference. The presence of ADAMTS-13 in platelets may imply the functional role of this enzyme in the local regulation of platelet function at the site of vascular injury or thrombus formation, and provide a useful tool for the analysis of structure and function of this enzyme.
Keywords:Platelets   vWF-CP   ADAMTS-13   vWF   TTP   Thrombosis   Expression   Real-time PCR
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