Structural bases of ADAMTS-13 and VWF A2 interactions

项目总结 这项研究将主要在#年在中国科学院生物物理研究所进行 北京，中国与楼继忠教授合作，作为美国国立卫生研究院R01 HL093723- 01A1。FIRCA的申请有一个应有的目标：1)扩大和加强父母赠款和2) 增加楼继忠博士团队的研究能力。这项提案和父提案的目标是 阐明von Willebrand因子与ADAMTS分子相互作用的机械调节 13，它们是生理性止血和病理性血栓形成的关键相互作用。机身出现故障 相互作用可能导致von Willebrand病和血栓性血小板减少性紫癜。ADAMTS- 13/VWF相互作用是机械调节的，因为它们发生在 发行量。我们对FIRCA赠款的假设是流体动力诱导VWF A2的展开 促进其与ADAMTS-13相互作用的结构域，与ADAMTS-13的结合也将导致 ADAMTS-13的构象变化对蛋白质的有效降解，构象受 A2展开程度和机械力。这一宽泛的假设将在两个具体目标中进行检验：1) 通过比较力和热诱导的去折叠来阐明2A型VWD的结构机制 野生型(WT)VWF A2结构域和一组突变的VWF A2结构域的途径，以及2)发展 ADAMTS-13/A2相互作用的原子模型并确定这些相互作用如何调节A2 展开和ADAMTS-13构象变化。这些具体目标是计算研究，它 补充家长助学金。分子动力学模拟和分子模拟 FIRCA赠款将帮助解释从父母赠款获得的实验数据，并提供 有机会设计新的实验，以检验父母和FIRCA赠款的总体假设。 破译机械力如何调节VWF A2的展开，ADAMTS-13的构象和 这两个分子之间的相互作用将为血管生理学和病理学提供关键的见解。AS 因此，这些数据可能会为相关疾病的治疗提供新的方法。

PROJECT SUMMARY This research will be done primarily at Institute of Biophysics, Chinese Academy of Sciences in Beijing, China in collaboration with Professor Jizhong Lou as an extension of NIH grant R01 HL093723- 01A1. This FIRCA application has a due objective: 1) to expand and enhance the parent grant and 2) to increase the research capacity of Dr. Jizhong Lou's group. The goal of this and the parent proposal is to elucidate the mechanical regulation of molecular interactions between von Willebrand factor and ADAMTS- 13, which are key interactions on physiological hemostasis and pathological thrombosis. Malfunction in the interactions may lead to von Willebrand disease and thrombotic thrombocytopenic purpura. ADAMTS- 13/VWF interaction are regulated mechanically as they take place in the hydrodynamic environment of the circulation. Our hypothesis for this FIRCA grant is that hydrodynamic forces induce the unfolding of VWF A2 domain which promote its interactions to ADAMTS-13, the binding with ADAMTS-13 will also lead to the conformational changes on ADAMTS-13 for efficient proteolysis and the conformations are regulated by the degree of A2 unfolding and mechanical forces. The broad hypothesis will be tested in two specific aims: 1) Elucidate the structural mechanisms of type 2A VWD by comparing the force- and thermo-induced unfolding pathways of wild-type (WT) VWF A2 domain and a panel of mutant VWF A2 domains, and 2) 2.Develop atomic models for the ADAMTS-13/A2 interactions and determine how these interactions regulate A2 unfolding and ADAMTS-13 conformational changes. These specific aims are computational studies, which complement the parent grant. The molecular dynamics simulations and molecular modeling proposed in the FIRCA grant will help to interpret the experimental data obtained from the parent grant and provide opportunities to design new experiments to test the overall hypothesis of the parent and the FIRCA grant. Decoding how mechanical forces regulate VWF A2 unfolding, ADAMTS-13 conformations and the interactions between the two molecules will provide key insights into vascular physiology and pathology. As a result, the data may office new therapeutic approaches to relevant diseases.