BLOOD COAGULATION FACTORS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 血液凝固是通过相互作用的促凝血因子和抗凝血因子之间的平衡来确保的。第VIII因子(FVIII)是一种大的多结构域蛋白(~280 kDa)，其活性形式为FVIIIa(FVIIIa)，与膜结合的Tenase复合体中的丝氨酸蛋白酶因子IXa(FixA)起辅助作用。 FVIIa与血小板表面FixA的结合使凝血因子Xa(FXA)和凝血酶的生成增加了106倍以上。FVIII基因突变导致轻微到严重的A型血友病，这是一种威胁生命的血液疾病，影响5000名男性人口中的一人(Wacey等人，1996年)。治疗这种情况的唯一方法是静脉注射FVIII，Stoilova-McPhie已经使用CryoEM研究了FVIII的膜结合结构。 我们的目标是确定血液凝血因子的膜结合复合体(ES)的活性构象(结构)，这些凝血因子对于止血的抗凝途径是必不可少的。我们的方法是将它们螺旋组织到一个脂质纳米管上，并定义它们的膜结合结构(结合)冷冻-EM和图像重建。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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 cryoEM. Our goal is to define the active conformation (structure) of the membrane-bound complex(es) of blood coagulation factors essential for the aniticoagulant pathways of blood hemostasis. Our approach is to organize them helically (onto) a lipid nanotube and define (their) membrane-bound structure (combining) Cryo-EM and image reconstruction.