STRUCTURE AND FUNCTION OF CASPASES

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 我们目前的工作重点是 caspase-6 和 -7。在本报告期内，我们致力于了解 caspase-6 和 -7 的五个主要目标。首先，我们设计了两类新的活性位点抑制剂。我们拥有带有多种活性位点抑制剂的 caspase-6 晶体。其次，我们设计了几种突变来探讨 caspase-6 中 130 和 90 螺旋结构变化的作用。第三，我们结晶了几种处于新失活状态的 caspase-6 版本。这些结构将帮助我们了解并最终控制神经退行性疾病中的 caspase-6。第四，我们生长了带有许多新型活性位点配体的 caspase-7 晶体。这些研究的目的是了解半胱天冬酶的反应机制。迄今为止，催化的分子细节仍然模糊，因为已进行结构表征的唯一活性位点配体也会扭曲催化二元组周围的几何形状。第五，我们有几个新的 caspase-7 突变体的晶体，旨在帮助我们了解 caspase-7 活性位点和变构位点调节的相互作用。

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. Our current work focuses on caspase-6 and -7. During this reporting period we worked on five main goals for understanding caspase-6 and -7. First, we have designed two new classes of active site inhibitors. We have crystals of caspase-6 with several of these active site inhibitors. Second, we have designed several mutations to probe the role of the structural changes in the 130's and 90's helix in caspase-6. Third, we have crystallized several versions of caspase-6 in a newly inactivated state. Together these structures will help us to understand and ultimately control caspase-6 in neurodegenerative diseases. Forth, we have grown crystals of caspase-7 with a number of novel active-site ligands. The aim of these studies is to understand the reaction mechanism in caspases. To date the molecular details of catalysis remain hazy because the only active site ligands that have been structurally characterized also distort the geometry around the catalytic diad. Fifth we have crystals of several new mutants of caspase-7 that are aimed at helping us to understand the interplay of caspase-7 active-site and allosteric-site regulation.