BLOOD COAGULATION FACTOR

凝血因子

• 批准号: 8168563
• 负责人: SVETLA STOILOVA-MCPHIE
• 金额: $ 2.15万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168563
• 关键词: Affect; Binding; Blood; Blood Coagulation Factor; Blood Platelets; Blood coagulation; Cells; Coagulants; Complex; Computer Retrieval of Information on Scientific Projects Database; Cryoelectron Microscopy; Crystallization; Electron Microscopy; Ensure; Environment; Equilibrium; Factor IXa; Factor VIII; Factor VIIIa; Factor Va; Factor Xa; Funding; Generations; Goals; Grant; Hemophilia A; Institution; Life; Lipids; Membrane; Methods; Mutation; Nanotubes; Population; Proteins; Research; Research Personnel; Resources; Serine Protease; Source; Structure; Surface; Thrombin; Time; United States National Institutes of Health; design; factor IXa-factor VIIIa; intravenous administration; male; protein structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Blood coagulation is ensured through equilibrium between pro- and anti-coagulant coagulation factors that interact with each other and cells. Factor VIII (FVIII) is a large multidomain protein (~280kDa), which in its active form, Factor VIIIa (FVIIIa) acts as a co-factor to the serine protease Factor IXa (FIXa) within the membrane-bound Tenase complex. Binding of FVIIIa to FIXa onto the platelet surface increases Factor Xa (FXa) and Thrombin generation more than 10^6 times. Mutations in FVIII result in mild to severe Haemophilia type A, a life-threatening blood condition affecting one in 5000 of the male population (Wacey et al., 1996). The sole cure for this condition is intravenous administration of FVIII, whose membrane-bound structure has been studied by Stoilova-McPhie using electron microscopy (Parmenter and Stoilova-McPhie, 2008; Stoilova-McPhie et al., 2008). CryoEM is a unique method capable of defining the structure of proteins in their native environment. For this purpose we have designed lipid nanotubes (LNT) for helical crystallization of FVIII and its complexes, allowing its membrane-bond structure to be resolved by Cryo-EM. Defining the structure of membrane-bound blood coagulation Factor Va and Factor VIIIa attached to lipid nanotubes by Cryo-electron microscopy is the primary goal of this project.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 通过促凝和抗凝凝血因子之间的平衡来确保血液凝固，这些凝血因子彼此之间以及细胞之间相互作用。因子 VIII (FVIII) 是一种大型多结构域蛋白 (~280kDa)，在其活性形式下，因子 VIIIa (FVIIIa) 在膜结合 Tenase 复合物中充当丝氨酸蛋白酶因子 IXa (FIXa) 的辅助因子。 FVIIIa 与 FIXa 结合到血小板表面可将 Xa 因子 (FXa) 和凝血酶的生成增加 10^6 倍以上。 FVIII 突变会导致轻度至重度 A 型血友病，这是一种危及生命的血液病，影响着五千分之一的男性人口（Wacey 等，1996）。这种情况的唯一治疗方法是静脉注射 FVIII，Stoilova-McPhie 使用电子显微镜研究了 FVIII 的膜结合结构（Parmenter 和 Stoilova-McPhie，2008；Stoilova-McPhie 等人，2008）。 CryoEM 是一种独特的方法，能够在天然环境中定义蛋白质的结构。为此，我们设计了用于 FVIII 及其复合物螺旋结晶的脂质纳米管 (LNT)，从而可以通过冷冻电镜解析其膜键结构。 通过冷冻电子显微镜定义附着在脂质纳米管上的膜结合凝血因子 Va 和因子 VIIIa 的结构是该项目的主要目标。