Molecular Basis of Blood Coagulation Regulation

凝血调节的分子基础

基本信息

批准号：
9230409
负责人：
Steven T. Olson
金额：
$ 39.18万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2020-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9230409
关键词：
Affinity Amino Acids Anticoagulants Antithrombins Bacteria Binding Binding Sites Biochemical Biological Assay Biotechnology Blocking Antibodies Blood Coagulation Disorders Blood coagulation Burial Cardiovascular Diseases Cause of Death Charge Coagulation Process Collaborations Communication Complex Coupling Disease Disulfides Down-Regulation Embryo Engineering Epitopes Equilibrium Factor IXa Factor Xa Fluorescence Generations Genes Hemophilia A Hemorrhage Hemostatic function Heparin Heparin Binding Human Hydrophobicity In Vitro Isotope Labeling Kinetics Knockout Mice Label Mediating Membrane Modeling Molecular Molecular Conformation Mus Mutagenesis Mutate Mutation Myocardial Infarction N-terminal Oxides Patients Peptide Hydrolases Pharmacologic Substance Phenotype Physiological Plasma Protease Inhibitor Protein Family Proteins Regulation Reporter Reporting Risk Factors Role Scanning Serpin Superfamily Serpins Signal Transduction Site Site-Directed Mutagenesis Sodium Chloride Stroke Tail Testing Thermodynamics Thrombin Thromboplastin Time Universities Up-Regulation Variant base biophysical techniques cancer procoagulant cofactor experimental study factor V Leiden fluorophore gain of function high throughput screening human subject instrument knockout animal novel strategies novel therapeutics oxidation plasma protein Z scaffold small molecule small molecule libraries transmission process virtual

项目摘要

Serpin family protein protease inhibitors function as key anticoagulant regulators of blood coagulation proteases. Two serpins, antithrombin and protein Z-dependent protease inhibitor (ZPI), are known to inhibit procoagulant proteases in a manner that is regulated by cofactors and dependent on the functional state of the proteases, but the molecular details of this complex regulatory mechanism are poorly understood. The physiologic importance of these serpins in regulating coagulation proteases is borne out by the observations that knocking out the mouse genes results in embryonic lethality due to a consumptive coagulopathy in the case of antithrombin and a prothrombotic phenotype when mice are bred on a factor V Leiden background in the case of ZPI. Our proposed studies seek to build on our prior studies to advance molecular understanding of the cofactor-dependent regulation of blood coagulation proteases by these serpins. With respect to antithrombin, our recent studies have suggested an important revision of the allosteric mechanism of activation of this serpin by heparin in showing that activation is mediated principally by the mitigation of repulsive interactions with factor Xa and factor IXa within an antithrombin exosite rather than by provision of an attractive exosite. Our proposed continuation studies seek to characterize the molecular determinants in antithrombin of protease binding exosites and of the allosteric communication network involved in transmitting activating conformational changes from the heparin binding site to the protease binding site. With respect to ZPI, our studies seek to elucidate the physiologic complexes of factor Xa that are targeted by ZPI and its cofactor, protein Z, establish the role of the unique N-terminal tail of ZPI in anticoagulant regulation of factor Xa and discover small molecules that disrupt the ZPI-protein Z complex and downregulate the ZPI anticoagulant mechanism as a potential novel therapy for restoring hemostasis in hemophilia bleeding disorders. Gain of function studies in which the serpin scaffold of α1-protease inhibitor is used to graft the molecular determinants of the protein Z binding site and protease binding sites onto ZPI will provide a stringent test of the minimal determinants of cofactor and protease recognition by this serpin.

SERPIN家族蛋白蛋白抑制剂充当血液凝血蛋白的关键抗凝剂调节剂。已知两种SERPINS抗凝血酶和蛋白Z依赖性蛋白抑制剂（ZPI）以由辅助因子调节的方式抑制proc凝蛋白，并依赖于蛋白质的功能状态，但是该复杂调节机制的分子细节知之甚少。这些SERPINS在控制凝结蛋白中的生理重要性是由观察结果出生的，即在抗强化蛋白的情况下，在抗强化抗强化的情况下，敲除小鼠基因而导致胚胎致死性，而当小鼠在Zpi的情况下，小鼠在因子V leiden的背景上繁殖小鼠时会导致胚胎致死性。我们提出的研究旨在以我们先前的研究为基础，以提高对这些Serpins对血液凝血蛋白酶的依赖性调节的分子理解。关于抗凝血酶，我们最近的研究表明，肝素通过肝素激活这种SERPIN的变构机制的重要修订是，表明激活主要是通过与因子XA和因子IXA在抗氧化抗凝结物中的反应性相互作用而介导的，而不是通过抗氧化抗合物蛋白外的exosite exosite介导的，而不是通过提供有吸引力的Exosite的替代性相互作用。我们提出的延续研究旨在表征蛋白质结合外os岩的抗凝结中的分子决定剂以及涉及将激活构象变化从肝素结合位点传输到蛋白质结合位点的变形构象变化的变构通信网络的分子决定剂。 With respect to ZPI, our studies seek to elucidate the physiological complexes of factor Xa that are targeted by ZPI and its cofactor, protein Z, establish the role of the unique N-terminal tail of ZPI in anticoagulant regulation of factor Xa and discover small molecules that disrupt the ZPI-protein Z complex and downregulate the ZPI anticoagulant mechanism as a potential novel therapy for restoring hemostasis in血友病出血疾病。功能研究的增益，其中使用α1-蛋白酶抑制剂的SERPIN支架嫁接蛋白质Z结合位点的分子确定剂，而蛋白质结合位点在ZPI上将提供严格的测试，以实现该蛇素识别的辅助因素和蛋白质识别的最小决定剂。