STRUCTURAL STUDIES OF HSP90 CHAPERONE-CLIENT INTERACTIONS

HSP90 伴侣-客户相互作用的结构研究

• 批准号: 8171520
• 负责人: DANIEL T GEWIRTH
• 金额: $ 0.71万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171520
• 关键词: Client; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Development; Funding; GRP94; Grant; Growth and Development function; Heat-Shock Proteins 90; Home environment; Institution; Ligand Binding Domain; Ligands; Malignant Neoplasms; Molecular Chaperones; N-terminal; Nature; Play; Proteins; Research; Research Personnel; Resolution; Resources; Roentgen Rays; Role; Source; Specificity; Staging; Structure; Time; United States National Institutes of Health; cell growth; novel; protein degradation; small molecule; synchrotron radiation; therapeutic target

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Hsp90 chaperones play central roles in the later stages of client protein maturation and, because these clients moderate key checkpoints in cellular growth and development, are recognized as significant cancer therapeutic targets. Questions at the forefront of the field currently center on the development of inhibitory ligands of high specificity, and the nature of the interaction between the chaperone and its client proteins and accessory factors. We are requesting beam time to study the interaction of the hsp90 family of molecular chaperones with novel small molecule inhibitory ligands and previously uncharacterized macromolecular partners. We have previously determined high resolution crystal structures of intact GRP94 (the endoplasmic recticulum Hsp90), as well as the N- terminal ligand binding domain. Crystals of the N-terminal domain in complex with inhibitory ligands typically diffract to better than 2 ¿ using synchrotron radiation. Crystals of the intact chaperone diffract to roughly 2.5 ¿ under favorable conditions. Complexes between the chaperone and macromolecular partners such as the Erad (ER associated degradation) protein are expected to diffract weakly and, because of their small size, are difficult to characterize using home X-ray sources. Our plan is to use CHESS beam time to 1) collect high resolution data from crystals of GRP94-Nterm plus inhibitory ligands, and 2) characterize and collect data from crystals of intact GRP94 in complex with the Erad protein.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Hsp90 伴侣在客户蛋白成熟的后期发挥核心作用，并且由于这些客户调节细胞生长和发育中的关键检查点，因此被认为是重要的癌症治疗靶点。目前该领域最前沿的问题集中在高特异性抑制性配体的开发，以及伴侣与其客户蛋白和辅助因子之间相互作用的性质。 我们正在请求光束时间来研究 hsp90 分子伴侣家族与新型小分子抑制配体和先前未表征的大分子伴侣的相互作用。我们之前已经确定了完整 GRP94（内质网 Hsp90）以及 N 端配体结合域的高分辨率晶体结构。使用同步加速器辐射，与抑制性配体复合的 N 端结构域晶体的衍射角度通常优于 2 ¡在有利的条件下，完整分子伴侣的晶体衍射大约为 2.5 ¿分子伴侣和大分子伴侣（例如 Erad（ER 相关降解）蛋白）之间的复合物预计衍射较弱，并且由于其尺寸小，很难使用家用 X 射线源来表征。我们的计划是使用 CHESS 光束时间 1) 从 GRP94-Nterm 加抑制配体的晶体收集高分辨率数据，以及 2) 从与 Erad 蛋白复合的完整 GRP94 晶体表征和收集数据。