STRUCTURAL STUDIES OF HSP90 CHAPERONE-CLIENT INTERACTIONS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 Hsp 90分子伴侣在晚期阶段的客户端中发挥核心作用 蛋白质成熟，因为这些客户温和的关键， 细胞生长和发育的检查点，被认为是 重要的癌症治疗靶点。最前沿的问题 目前，该领域的研究主要集中在抑制性药物的开发上。 高特异性的配体，以及相互作用的性质 分子伴侣与其客户蛋白和辅助因子之间的关系。 我们正在申请光束时间来研究热休克蛋白90 具有新的小分子抑制剂的分子伴侣家族 配体和以前未表征的大分子伴侣。我们 先前已经确定了高分辨率的晶体结构， 完整的GRP 94（内质网Hsp 90），以及N- 末端配体结合域。晶体的N-末端结构域， 与抑制性配体的复合物通常作用于优于2？ 使用同步辐射。完整分子伴侣的晶体 在有利的条件下，它可以达到大约2.5美分。配合物 分子伴侣和大分子伴侣之间， (ER相关降解）蛋白的降解作用较弱 并且由于它们的尺寸小，难以使用 家用X射线源我们的计划是利用国际象棋光束时间1）收集 来自GRP 94-Nterm加抑制剂晶体的高分辨率数据 配体，和2）表征和收集数据，从晶体的完整的 GRP 94与Erad蛋白复合。

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.