Structural analysis of oxidation effects on the hemostatic process

氧化对止血过程影响的结构分析

• 批准号: 8486261
• 负责人: Gianluca Interlandi
• 金额: $ 11.15万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-02 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8486261
• 关键词: Adhesions; Adhesiveness; Affect; Area; Award; Binding; Biological Assay; Blood Platelets; Clinical Pathology; Coagulation Process; Complement; Cytolysis; Data; Disease; Drug Design; Environment; Enzymes; Equilibrium; Equipment; Faculty; Fiber; Fibrin; Fibrinogen; Goals; Hemorrhage; Hemostatic Agents; Hemostatic function; Inflammation; Knowledge; Laboratories; Lateral; Lead; Learning; Life; Light; Link; Literature; Location; Manuscripts; Mass Spectrum Analysis; Mechanics; Mentors; Mentorship; Methionine; Methods; Microscopy; Modification; Molecular; Molecular Structure; Mutagenesis; Mutation; Oxidants; Pathologic; Pathology; Pathway interactions; Patients; Plasma; Platelet aggregation; Positioning Attribute; Process; Production; Property; Proteins; Reporting; Research; Resources; Risk; Running; Site; Structure; Supercomputing; Testing; Therapeutic; Thrombin; Thrombosis; Thrombotic Thrombocytopenic Purpura; Thrombus; Universities; Washington; base; career; design; drug discovery; insight; methionine sulfoxide; molecular dynamics; oxidation; polymerization; prevent; protein structure; public health relevance; research study; simulation; skills; structural biology; von Willebrand Factor

DESCRIPTION (provided by applicant): The long-term career goal of the candidate is to investigate the structural basis of hemostasis and how external factors alter it. During the K25 award he will analyze how oxidation effects alter the structure and function of two key proteins in the hemostatic process, von Willebrand Factor and fibrin. The candidate will use molecular dynamics simulations, his main expertise, to investigate how oxidation alters the structural stability of proteins. Dr. Jim Pfaendtner, an expert in accelerated molecular dynamics methods, will serve as a co-mentor. The simulations will be complemented through microscopy experiments which the candidate will learn how to perform himself under the mentorship of Dr. Wendy Thomas. Thanks to Dr. Jose Lopez as co-mentor, the candidate will gain deep knowledge of pathologies related to hemostasis. In conclusion, the candidate will learn exceptional skills outside the area of his primary expertise in order to become an essential link between clinical pathologies and structural biology. Environment. The University of Washington has a strong track record of strength in both computational protein structure analysis and in hemostasis and thrombosis research. During the transition to an independent position the candidate will be mentored by Dr. Thomas, Dr. Pfaendtner and Dr. Lopez. He will also collaborate with various other faculties like Dr. Nathan White and Dr. Xiaoyun Fu. Dr. Thomas will provide him with the necessary computational and experimental equipment and office space also after his transition to independence. The computations will be run mainly on national supercomputing resources while the experiments will be performed in the laboratory of Dr. Thomas or in the laboratories of collaborating faculties. Description. Hemostasis is an essential mechanism to prevent blood loss from a damaged vessel. It is a complicated pathway that can become out of balance and lead for example to the formation of life threatening thrombi. Inflammation leads to the production of oxidizing agents. It has been reported in the literature that oxidation alters the structure of fibrin clot and influences the function of von Wille- brand Factor. However, it is currently not understood how the molecular structure of fibrin molecules and domains of von Willebrand Factor is altered due to oxidation in order to explain the experimentally observed functional modifications. The research proposed here investigates the link between altered fibrin clot structure and structural changes in single fibrin molecules, and the altered function of von Willebrand Factor as the result of oxidation. For this purpose, a combination of molecular dynamics simulations and flow chamber experiments will be used. Understanding the function of VWF and the process of fibrin polymerization under oxidizing conditions will help the design of therapeutics that can be administered to patients who are at risk of thrombosis due to inflammation.

描述(由申请人提供)：应聘者的长期职业目标是调查止血的结构基础以及外部因素如何改变它。在K25奖项中，他将分析氧化效应如何改变止血过程中两种关键蛋白质的结构和功能，这两种蛋白质是von Willebrand因子和纤维蛋白。候选人将使用分子动力学模拟，这是他的主要专长，来研究氧化如何改变蛋白质的结构稳定性。加速分子动力学方法专家吉姆·普芬特纳博士将担任共同导师。模拟实验将通过显微镜实验来补充，候选人将在温迪·托马斯博士的指导下学习如何表现自己。多亏了何塞·洛佩兹博士的共同指导，候选人将获得与止血相关的病理学方面的深入知识。总之，候选人将学习他的主要专业领域之外的特殊技能，以便成为临床病理学和结构生物学之间的重要纽带。环境华盛顿大学在计算蛋白质结构分析以及止血和血栓研究方面都有很强的实力。在过渡到独立职位的过程中，候选人将得到托马斯博士、普芬特纳博士和洛佩兹博士的指导。他还将与内森·怀特博士和傅晓云博士等其他院系合作。托马斯博士也将在他过渡到独立后为他提供必要的计算和实验设备以及办公空间。计算将主要在国家超级计算资源上运行，而实验将在托马斯博士的实验室或合作学院的实验室进行。描述。止血是防止受损血管失血的重要机制。这是一个复杂的途径，可以变得失衡，并导致例如威胁生命的血栓的形成。炎症会导致氧化剂的产生。已有文献报道，氧化改变纤维蛋白凝块的结构，影响von Wille-brand因子的功能。然而，为了解释实验观察到的功能修饰，目前还不清楚von Willebrand因子的纤维蛋白分子和结构域的分子结构是如何因氧化而改变的。这项研究旨在研究纤维蛋白凝块结构改变与单个纤维蛋白分子结构改变之间的联系，以及von Willebrand因子在氧化过程中的功能改变。为此，将使用分子动力学模拟和流室实验相结合的方法。了解VWF的功能和氧化条件下纤维蛋白聚合的过程将有助于设计可用于因炎症而有血栓形成风险的患者的治疗方法。