UNDERSTANDING THE ARCHITECTURE OF CHALLENGING MULTI-DOMAIN PROTEINS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 这是一个描述我们实验室正在研究的多结构域和多蛋白质组装的研究建议。Gulick实验室正在研究非核糖体肽合成酶（NRPS）的结构排列。这些模块化蛋白质使用与载体结构域共价结合的氨基酸和肽中间体催化肽抗生素和肽铁载体的合成，所述载体结构域以协调的方式将底物递送至必要的催化结构域。这种装配线合成的编排正在研究通过生物化学研究和结构研究的多结构域蛋白质被困在催化相关的状态。其他项目研究在天然产物合成中与NRPS酶起作用的辅助蛋白。Gebruth实验室研究了多结构域HSP 90类分子伴侣。目前，该领域最前沿的问题集中在开发高特异性的抑制性配体，以及了解细胞质Hsp 90和GRP 94之间的机制差异。我们正在使用x射线晶体学结合生化和细胞为基础的分析，研究热休克蛋白90家族的分子伴侣与新的小分子抑制配体的相互作用。

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. This is a proposal describing research into multi-domain and multi-protein assemblies under study in our labs. The Gulick lab is investigating the structural arrangement of non-ribosomal peptide synthetases (NRPSs). These modular proteins catalyze the synthesis of peptides antibiotics and peptide siderophores using amino acid and peptide intermediates that are covalently bound to a carrier domain that delivers substrates to the necessary catalytic domains in a coordinated fashion. The choreography of this assembly line synthesis is being investigated through biochemical studies and structural investigations of multi-domain proteins trapped in catalytically relevant states. Additional projects investigate auxiliary proteins that function with NRPS enzymes in the synthesis of natural products. The Gewirth lab studies the multi-domain HSP90 class of molecular chaperones. Questions at the forefront of the field currently center on the development of inhibitory ligands of high specificity, as well as understanding the mechanistic differences between cytoplasmic Hsp90 and GRP94. We are using x-ray crystallography in combination with biochemical and cell-based assays to study the interaction of the hsp90 family of molecular chaperones with novel small molecule inhibitory ligands.