Hemostasis and Thrombosis: Chemistry, Biology and Physiology

止血和血栓形成：化学、生物学和生理学

• 批准号: 9982398
• 负责人: Sriram Krishnaswamy
• 金额: $ 206.71万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 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/9982398
• 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; 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; prothrombinase complex; response; structural biology

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.

总体计划-摘要 凝血源于一系列特定的蛋白水解性激活反应，这些反应是由 胰酶样丝氨酸蛋白酶的狭义和明确的特异性。在几种情况下，这些蛋白酶 在膜组装酶复合体中的作用。独特的蛋白质底物特异性和 通过与膜和辅因子的相互作用来调节酶的功能是蛋白质分解的标志 凝血反应。目前对分子碱基的理解存在很大差距 这些独特的特征构成了止血反应的功能基础。该计划提出了一种 综合方法侧重于调节酶的功能和特异性，其独特之处在于 作为止血酶作用基础的大分子相互作用。项目1(Krishnaswamy) 使用凝血酶原酶复合体作为范例来研究结构和功能机制 从辅因子、蛋白酶和底物在酶功能中的膜依赖性相互作用。 项目2(CAMIRE)将研究因子V转化为辅因子的分子机制， 因子Va和这些机制揭示的令人惊讶的新生物学见解。项目3(Sullener) 使用RNA适配子作为独特的探针，用于研究凝血所必需的大分子相互作用 酶的功能，并指明了干扰或增强这些功能的新方法的前进方向 相互作用以获得治疗收益。项目4(Arruda)重点研究内在Xase的生物化学 具有优越功能和下一代应用潜力的蛋白酶和辅因子突变体的鉴定 治疗血友病的新一代疗法。四个项目的目标将得到支持 由管理核心(核心A)提供分子生物学、蛋白质表达和 结构生物学(核心B)和一个核心，该核心支持该计划的小鼠止血和 血栓形成(核心C)。总体而言，该项目将个别调查人员的专门知识应用于 解决止血和血栓形成方面的主要悬而未决的问题，从生化见解扩展到 生物功能和生理学。建议的方法将提供对化学和化学的新见解 凝血反应的生物学及其对理解正常止血和 血栓形成。