Hemostasis and Thrombosis: Chemistry, Biology and Physiology

止血和血栓形成：化学、生物学和生理学

基本信息

批准号：
9769852
负责人：
Sriram Krishnaswamy
金额：
$ 254.87万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9769852
关键词：
Address Area Biochemical Biochemistry Biological Biological Process Biology Blood Vessels Blood coagulation Chemistry Coagulation Process Complex Deuterium Disease Ensure Enzymes Factor V Factor Va Foundations Goals Hemophilia A Hemorrhage Hemostatic Agents Hemostatic function Hydrogen Individual Kinetics Knowledge Lead Life Membrane Membrane Proteins Molecular Molecular Biology Multienzyme Complexes Myocardial Infarction Participant Peptide Hydrolases Physiology Play Proteins RNA Reaction Reagent Recombinants Records Regulation of Proteolysis Research Research Personnel Roentgen Rays Role Series Serine Protease Specificity Stroke Structural Protein Structure Substrate Interaction Substrate Specificity Testing Therapeutic Thrombosis Thrombus Trypsin Variant Western World X-Ray Crystallography aptamer base biophysical analysis biophysical techniques cancer procoagulant cofactor enzyme substrate human disease human morbidity human mortality insight interdisciplinary approach interest mouse model novel strategies novel therapeutics programs protein expression protein structure prothrombinase complex response structural biology

项目摘要

Overall Program-Abstract Blood coagulation derives from a series of specific proteolytic activation reactions that are catalyzed with narrow and defined specificity by trypsin-like serine proteinases. In several instances, these proteinases function in membrane assembled enzyme complexes. Distinctive protein substrate specificities and the modulation of enzymic function by interactions with membranes and cofactors are hallmarks of the proteolytic reactions of blood coagulation. There are major gaps in the current understanding of the molecular bases for these unique features that underlie the function of the hemostatic reactions. This program proposes an integrated approach focused on the modulation of enzymic function and specificity that uniquely arises from macromolecular interactions that underlie the action of the hemostatic enzymes. Project 1 (Krishnaswamy) uses the prothrombinase complex as a paradigm to investigate structural and functional mechanisms deriving from the membrane-dependent interactions between cofactor, proteinase and substrate in enzyme function. Project 2 (Camire) will investigate molecular mechanisms at play in the conversion of factor V to the cofactor, factor Va and the surprising new biological insights that these mechanisms reveal. Project 3 (Sullenger) employs RNA aptamers as unique probes for the macromolecular interactions essential for coagulation enzyme function and to show the way forward for novel approaches to either interfere with or enhance these interactions for therapeutic gain. Project 4 (Arruda) investigates the biochemistry of intrinsic Xase with a focus on characterizing proteinase and cofactor variants with superior function and potential for use as the next generation of therapeutics in the treatment of hemophilia. The objectives of the four projects will be supported by an administrative core (Core A), a core that provides support for molecular biology, protein expression and structural biology (Core B) and a core that supports the program with mouse models of hemostasis and thrombosis (Core C). Overall, this project applies the expertise of the individual investigators towards addressing major unanswered questions in hemostasis and thrombosis extending from biochemical insights to biological function and physiology. 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 thrombosis.

总体程序提取血液凝血来自一系列特定的蛋白水解激活反应，这些反应与通过胰蛋白酶样丝氨酸蛋白酶狭窄而定义的特异性。在某些情况下，这些蛋白酶膜组装酶复合物中的功能。独特的蛋白质底物特异性和通过与膜和辅因子相互作用的酶功能调节是蛋白水解的标志血液凝血的反应。当前对分子碱基的理解存在主要差距这些独特的特征是止血反应功能的基础。该程序提出了一个集成方法着重于酶功能和特异性的调节止血酶作用的基础的大分子相互作用。项目1（克里希纳斯瓦米）使用凝血酶酶复合物作为范式来研究得出的结构和功能机制从酶功能中的辅因子，蛋白酶和底物之间的膜依赖性相互作用。项目2（Camire）将研究因子V向辅因子转化时的分子机制，因子VA和这些机制所揭示的令人惊讶的新生物学见解。项目3（Sullenger）利用RNA适体作为对凝结必不可少的大分子相互作用的独特探针酶功能并展示新方法干扰或增强这些方法的前进方向治疗增益的相互作用。项目4（Arruda）研究了内在Xase的生物化学，重点在表征具有优质功能且潜在使用的蛋白酶和辅因子变体作为下一个治疗血友病的治疗剂。这四个项目的目标将得到支持通过行政核心（核心A），该核心为分子生物学，蛋白质表达和结构生物学（核心B）和核心止血模型和核心的核心血栓形成（核心C）。总体而言，该项目将个人调查人员的专业知识应用于解决止血和血栓形成中的主要未解决问题，从生化见解延伸到生物功能和生理。拟议的方法将为化学和血液凝血反应的生物学，对正常止血和血栓形成。