LIFETIME IMAGING OF COAGULATION ENZYME COMPLEXES

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。 凝血的许多蛋白水解性反应是由膜结合的大分子酶复合体催化的，该复合体由丝氨酸蛋白酶和辅因子蛋白组成。我们已经开发了一系列荧光衍生品，可以对辅因子-蛋白酶和底物-酶复合体的相互作用进行FRET测量。我们的目标是扩展这些研究，以允许在活着的小鼠体内进行酶复合体组装和底物在血细胞上的传递以及最终在血管系统中的寿命成像。我们想做一系列实验来评估这种方法的可行性，使用支撑的膜双层和/或培养的贴壁细胞。

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. Many of the proteolytic reactions of blood coagulation are catalyzed by membrane-bound macromolecular enzyme complexes composed of a serine proteinase associated with a cofactor protein. We have developed a series of fluorescent derivatives that allow FRET measurements of the cofactor-proteinase and substrate-enzyme complex interactions. Our goal is to extend these studies to permit lifetime imaging on enzyme complex assembly and substrate delivery on blood cells and eventually in the vasculature in the living mouse. We would like to do a series of experiments to assess the feasibility of such approaches using supported membrane bilayers and/or cultured adherent cells.