CONFORMATIONAL CHANGES AND MODE OF BINDING OF AGONIST TO ESTROGEN RECEPTOR

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 David Budil要求我们用电子自旋标记技术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反应。最初阶段的技术目标包括开发ER-LBD的定点自旋标记突变体，并为拟议的研究合成新的自旋标记配体。Budil的研究小组优化了ER-LBD的细胞培养生产，并表达了一些单标记和双标记突变体。此外，还合成了一种新的自旋标记，用于连接该蛋白质中选择位点的半胱氨酸残基，并在合成具有一系列激动剂/拮抗剂活性的自旋标记配体方面取得了实质性进展。初步的电子自旋共振研究已经确定，所选择的标记位置对存在的配体类型将是可接受的敏感的。

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.