Physiological and biochemical analysis of VWF-cleaving protease ADAMTS13

血浆ADAMTS13裂解von Willebrand因子(VWF)多聚体，调节其在血小板聚集中的活性。ADAMTS13缺乏会导致血栓性血小板减少性紫癜(TTP)。这项研究的目的是通过对ADAMTS13的基本分析，产生临床有用的成果。[ADAMTS13缺陷小鼠分析]产生缺失ADAMTS13基因的小鼠。他们表现出血栓前状态，但没有自发性血小板减少、溶血性贫血和微血管血栓形成的症状。这表明，小鼠完全缺乏ADAMTS13不足以引起TTP，除ADAMTS13缺乏外，其他因素可能是TTP发生所必需的。[携带截短的ADAMTS13的同源小鼠的产生与分析]我们先前发现了两种小鼠ADAMTS13：129/Sv株ADAMTS13与人的ADAMTS13具有相同的结构域，C57BL/6株ADAMTS13缺乏C端多个结构域。为了评估这些结构域在体内的意义，我们创建并分析了携带C57BL/6型ADAMTS13的129/Sv遗传背景同基因小鼠。[生化和流行病学分析]采用基于荧光的分析方法，对ADAMTS13的生化特征进行分析。在日本人群中经常观察到的ADAMTS13的P475S形式显示出大约70%的野生型活性。[寻找与ADAMTS13相互作用的蛋白]我们用酵母双杂交系统鉴定了与ADAMTS13相互作用的候选蛋白，其中一种被证实与ADAMTS13直接结合。本研究提供了有关ADAMTS13的新信息，这将对即将到来的翻译研究有所帮助。

Plasma ADAMTS13 cleaves von Willebrand factor (VWF) multimers to regulate their activity in platelet aggregation. ADAMTS13 deficiency results in thrombotic thrombocytopenic purpura (TTP). The aim of this research is the production of clinically useful achievement through the basic analysis of ADAMTS13. [Analysis of ADAMTS13-deficient mice] Mice lacking the Adamts13 gene were generated. They showed a prothrombotic state but no symptoms of spontaneous thrombocytopenia, hemolytic anemia, and microvascular thrombosis. This suggested that a complete lack of ADAMTS13 in mice was not sufficient to cause TTP and other factors in addition to ADAMTS13 deficiency may be necessary for development of TTP. [Generation and analysis of congenic mice carrying truncated ADAMTS13] We previously identified two kinds of mouse ADAMTS13:129/Sv-strain ADAMTS13 with the same domains as human ADAMTS13 and C57BL/6-strain ADAMTS13 lacking the C-terminal multiple domains. To assess the significance of these domains in vivo, we generated and analyzed 129/Sv-genetic-background congenic mice that carry the C57BL/6-type ADAMTS13. [Biochemical and epidemiological analysis] Using a fluorescence-based assay, the biochemical features of ADAMTS13 were analyzed. The P475S form of ADAMTS13, frequently observed in the Japanese population, showed an approximately 70% activity of the wild type. [Search of ADAMTS13-interacting proteins] We used a yeast two-hybrid system to identify the candidates of ADAMTS13-interacting proteins, one of which was confirmed to bind directly to ADAMTS13. The present study provided new information about ADAMTS13, which will be useful for oncoming translational research.