Structure and Function of Protein-Membrane Interactions in Blood Clotting

血液凝固中蛋白质-膜相互作用的结构和功能

• 批准号: 8644862
• 负责人: James H. Morrissey
• 金额: $ 53.29万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-06 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8644862
• 关键词: Affinity; Binding; Binding Sites; Blood Coagulation Factor; Blood coagulation; Calcium ion; Cessation of life; Choline; Coagulation Process; Complex; DNA Sequence Rearrangement; Dependence; Disease; Environment; Enzymes; Ethanolamines; Extracellular Domain; Factor VIIa; Factor X; Health; Label; Lecithin; Ligands; Light; Lipids; Magic; Measurement; Membrane; Membrane Proteins; Molecular; Molecular Conformation; Mutagenesis; Phosphatidic Acid; Phosphatidylethanolamine; Phosphatidylglycerols; Phosphatidylinositols; Phosphatidylserines; Phospholipid Interaction; Phospholipids; Phosphoserine; Process; Protein Binding; Proteins; Reaction; Recombinants; Regulation; Resolution; Role; Scaffolding Protein; Serine; Specificity; Sphingomyelins; Structure; Surface; Surface Plasmon Resonance; System; Techniques; Technology; Therapeutic Intervention; Thromboplastin; United States; Vesicle; Writing; disability; inorganic phosphate; insight; molecular dynamics; nanoscale; novel; protein structure function; public health relevance; restraint; simulation; solid state nuclear magnetic resonance

DESCRIPTION (provided by applicant): Almost every step in blood coagulation requires assembly of multiple proteins on membrane surfaces, yet protein-membrane interactions in blood clotting remain poorly understood at the molecular level. We are using new, high-resolution technologies including magic-angle spinning solid-state NMR (SSNMR) to probe the mechanisms by which blood clotting proteins interact with phospholipid surfaces, and how changes in membrane composition regulate clotting reactions. The primary focus of our studies is the membrane-bound complex of tissue factor and factor VIIa, the two-subunit enzyme responsible for triggering blood clotting in health and disease. Aim 1 will delineate, at atomic resolution, the changes in structure and dynamics of PS- rich membrane domains in the presence of Ca2+, and will examine the roles of phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine in configuring the membrane to support high affinity binding of clotting factors. Aim 2 will investigate the role of phospholipid-phospholipid interactions in enhancing blood clotting reactions on membrane surfaces. Aim 3 will delineate the lipid environments/conformations induced when clotting proteins bind to bilayers. Aim 4 will solve the structure of tissue factor on the membrane surface and identify conformational changes in tissue factor when it interacts with ligands. Together these studies will provide valuable new insights into the role of the membrane surface in blood clotting, at atomic-scale resolution. PUBLIC HEALTH RELEVANCE: Disorders of the blood clotting system represent the leading cause of disability and death in the United States, but we still have a very incomplete understanding of blood clotting reactions at the molecular level. These studies will shed new light on the regulation of the blood clotting system, with a particular focus on achieving a detailed understanding of how and why blood clotting reactions occur on membrane surfaces.

描述（由申请人提供）：几乎每个步骤的血液凝血都需要在膜表面组装多种蛋白质，但是在分子水平上，血液凝结中的蛋白质 - 膜相互作用仍然很少了解。我们正在使用新的高分辨率技术，包括魔法旋转固态NMR（SSNMR）来探测血液凝结蛋白与磷脂表面相互作用的机制，以及膜组成的变化如何调节凝血反应。我们研究的主要重点是组织因子和因子VIIA的膜结合的复合物，这是负责触发健康和疾病血液凝血的两生酶。 AIM 1将以原子分辨率描绘出Ca2+存在的PS-富膜结构域的结构和动力学的变化，并将检查磷脂酰丝氨酸，磷脂酰胆碱和磷脂酰乙醇胺在配置膜以支持凝结因子的高亲和力结合的膜上的作用。 AIM 2将研究磷脂 - 磷脂相互作用在增强膜表面上的血液凝结反应中的作用。 AIM 3将描绘凝血蛋白与双层蛋白结合时引起的脂质环境/构象。 AIM 4将解决膜表面的组织因子的结构，并在与配体相互作用时确定组织因子的构象变化。这些研究将共同​​提供有价值的新见解，以解决原子级分辨率下膜表面在血液凝结中的作用。 公共卫生相关性：血液凝结系统的疾病代表了美国残疾和死亡的主要原因，但我们对分子水平上血液凝结反应的了解仍然非常不完整。这些研究将为血液凝结系统的调节提供新的启示，特别着重于对膜表面上的血液凝结反应的详细理解。

项目成果

31P-dephased, 13C-detected REDOR for NMR crystallography at natural isotopic abundance.

31P 相移、13C 检测的 REDOR，用于天然同位素丰度的 NMR 晶体学。

• DOI: 10.1016/j.jmr.2017.02.020
• 发表时间: 2017
• 期刊: Journal of magnetic resonance (San Diego, Calif. : 1997)
• 作者: Greenwood,AlexanderI;Clay,MaryC;Rienstra,ChadM
• 通讯作者: Rienstra,ChadM

Recent advances in solid-state nuclear magnetic resonance techniques to quantify biomolecular dynamics.

量化生物分子动力学的固态核磁共振技术的最新进展。

• DOI: 10.1021/ac403956k
• 发表时间: 2014
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Watt,EricD;Rienstra,ChadM
• 通讯作者: Rienstra,ChadM

Influence of membrane composition on the enhancement of factor VIIa/tissue factor activity by magnesium ions.

膜组成对镁离子增强因子 VIIa/组织因子活性的影响。

• DOI: 10.1160/th13-07-0628
• 发表时间: 2014
• 期刊: Thrombosis and haemostasis
• 影响因子: 6.7
• 作者: Tavoosi,Narjes;Morrissey,JamesH
• 通讯作者: Morrissey,JamesH

High-Resolution NMR Studies of Human Tissue Factor.

• DOI: 10.1371/journal.pone.0163206
• 发表时间: 2016
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Nuzzio KM;Watt ED;Boettcher JM;Gajsiewicz JM;Morrissey JH;Rienstra CM
• 通讯作者: Rienstra CM