Chemistry and Biology of Coagulation

凝固化学与生物学

• 批准号: 6674562
• 负责人: Sriram Krishnaswamy
• 金额: $ 189.89万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-15 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6674562
• 关键词: blood coagulation; clinical research; clotting factor; human tissue; intermolecular interaction; protein structure function; serine proteinases

DESCRIPTION (provided by applicant): Blood coagulation derives from a series of specific proteolytic activation reactions that are catalyzed with narrow and defined specificity by trypsin-like serine proteinases. Many of these proteinases function in membrane assembled enzyme complexes. There are major gaps in the current understanding of the molecular bases for the distinctive protein substrate specificities and the modulation of enzymic function by interactions with membranes and cofactors that are hallmarks of the proteolytic reactions of blood coagulation. This program proposes an integrated approach to focus on the contributions of macromolecular interactions to specificity and the modulation of enzymic function to explain the action of the coagulation enzymes in vivo. Project 1 (Krishnaswamy) uses the prothrombinase complex as a paradigm to investigate the role of extended macromolecular interactions between the protein substrate and the enzyme complex in explaining protein substrate specificity and cofactor function. Project 2 (Camire) will investigate the relationship between procofactor and zymogen proteolysis and the development of discrete macromolecular binding interactions that lead to the assembly of the prothrombinase complex and the expression of enzymic function. Using the x-ray structure of the catalytic domain of XIa, Project 3 (Walsh) proposes structure function studies to examine the contributions of liganding interactions at extended macromolecular recognition surfaces to XIa function. These approaches in basic biochemistry and enzymology are complemented by Project 4 (High) that proposes to use genetic mouse models to investigate the roles of factors X and Xa in vivo and explore the relative contributions of Xa formation via the extrinsic and intrinsic pathways to hemostasis and thrombosis. The objectives of the four projects will be supported by an administrative core and a core that provides support for molecular biology and protein expression. Overall, this program applies the expertise of the individual investigators towards addressing major unanswered questions in the coagulation field. The proposed approaches will provide new insights into the chemistry and biology of the blood coagulation reactions with implications for an understanding of normal hemostasis and will suggest novel therapeutic strategies for targeting thrombosis and vascular disease.

描述（由申请人提供）：血液凝血源自一系列特定的蛋白水解活化反应，这些反应被胰蛋白酶样丝氨酸蛋白酶催化狭窄和定义的特异性。这些蛋白酶中的许多在膜组装的酶复合物中起作用。当前对分子碱基的特异性分子碱基的理解存在主要差距，以及通过与膜和辅助因子的相互作用来调节酶功能，这是血液凝结的蛋白水解反应的标志。该计划提出了一种集成的方法，以关注大分子相互作用对特异性的贡献以及酶功能的调节，以解释体内凝血酶的作用。 Project 1（Krishnaswamy）使用凝血酶酶复合物作为范式来研究蛋白质底物与酶复合物之间扩展的大分子相互作用在解释蛋白质底物特异性和辅因子功能中的作用。项目2（Camire）将研究profactor和Zymogen蛋白水解之间的关系，以及离散的大分子结合相互作用的发展，这些相互作用导致凝血酶酶复合物的组装和酶功能的表达。使用XIA催化结构域的X射线结构，项目3（WALSH）提出了结构功能研究，以检查扩展大分子识别表面在扩展的大分子识别表面对XIA功能的贡献。这些基本生物化学和酶学中的这些方法得到了项目4（高）的补充，这些方法提议使用遗传小鼠模型研究体内因子X和XA的作用，并探索XA形成通过外在和内在途径的相对贡献，这些途径的相对贡献。这四个项目的目标将由管理核心和核心支持，该核心为分子生物学和蛋白质表达提供支持。总体而言，该计划将个别调查人员的专业知识应用于解决凝血领域的主要未解决问题的专业知识。所提出的方法将提供有关血液凝血反应的化学和生物学的新见解，对了解正常止血的影响，并提出了针对血栓形成和血管疾病的新型治疗策略。