CONFORMATIONAL CHANGES AND MODE OF BINDING OF AGONIST TO ESTROGEN RECEPTOR

该子项目是利用 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来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 大卫Budil要求我们使用电子自旋标记来表征雌激素受体配体结合域（ER-LBD）对从强雌激素到强抗雌激素的各种配体的结构响应。初期的技术目标包括开发ER-LBD的定点自旋标记突变体和合成用于拟议研究的新自旋标记配体。Budil的研究小组已经优化了ER-LBD的细胞培养生产，并表达了许多单标记和双标记的突变体。此外，合成了一种新的自旋标记物，用于连接到该蛋白质中的位点选择性半胱氨酸残基，并且朝着具有一系列激动剂/拮抗剂活性的自旋标记配体的合成取得了实质性进展。最初的电子自旋共振研究已经确定，所选择的标记位置将是可接受的敏感的配体的类型存在。

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. David Budil requested us to characterize structural response of the estrogen receptor ligand binding domain (ER-LBD) to a variety of ligands ranging from strong estrogens to strong antiestrogens using electron spin labeling. The technical aims for the initial period involved developing site-directed spin-labeled mutants of the ER-LBD and synthesizing new spin-labeled ligands for the proposed studies. Budil's research group has optimized cell culture production of ER-LBD and expressed a number of single and doubly labeled mutants. In addition a new spin label was synthesized for attachment to site-selected cysteine residues in this protein and substantial progress was made towards the synthesis of spin-labeled ligands with a range of agonist/antagonist activities. Initial electron spin resonance studies have established that the selected label locations will be acceptably sensitive to the type of ligand present.