UNDERSTANDING THE ARCHITECTURE OF CHALLENGING MULTI-DOMAIN PROTEINS

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。 这是一项描述我们实验室正在研究的多域和多蛋白质组装研究的提案。古力克实验室正在研究非核糖体多肽合成酶(NRPS)的结构安排。这些模块化蛋白利用氨基酸和多肽中间体催化合成多肽、抗生素和多肽铁载体，这些中间体共价结合到载体结构域，该载体结构域以协调的方式将底物输送到必要的催化结构域。通过生化研究和对处于催化相关状态的多域蛋白质的结构研究，正在研究这种装配线合成的编排。其他项目研究在天然产物合成中与NRPS酶一起发挥作用的辅助蛋白质。Gewirth实验室研究多结构域HSP90类分子伴侣。目前该领域的前沿问题集中在高特异性抑制配体的开发，以及了解细胞质Hsp90和GRP94之间的机制差异。我们正在使用X射线结晶学结合生化和基于细胞的分析方法来研究HSP90分子伴侣家族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.