Folding of Androgen Receptor-Coregulator Complex

描述（由申请人提供）：雄激素受体是男性性激素睾酮的细胞介质，是前列腺癌发生和发展的关键调节因子。雄激素受体在核受体家族中是不同寻常的，其与激素无关的AF-1活性是显性基因的转激活功能。AF-1的活性已被映射到n端结构域的一个区域。这个结构域包含许多共激活因子、辅抑制因子和其他蛋白质的相互作用位点。尽管AF-1在雄激素受体功能和前列腺癌病理中起着关键作用，但AF-1活性的结构基础以及更普遍的n端雄激素受体结构域的功能尚不清楚。我们将定义、表达、纯化和结晶稳定的n端雄激素受体结构域片段配合50个共调节蛋白（如SRC和ARA）的代表性子集的结构域的复合物。我们的建议采用高通量机器人表达和纯化系统来筛选数千对n端受体结构域和共调节片段的组合，以发现稳定复合物的协同折叠结构域。雄激素受体-共调节剂复合物的结构将揭示设计第一个用于控制前列腺癌的n端受体区域的药物所必需的相互作用。通过影响n端雄激素受体结构域与类固醇受体共激活因子相互作用的药物拮抗雄激素受体活性的能力有望抑制前列腺癌的转移性侵袭和增殖。我们的研究目标是建立一种发现策略，通过破坏雄激素受体n端结构域和关键协同调节因子之间的关键相互作用，识别前列腺癌肿瘤中雄激素受体活性拮抗剂的小有机化合物。

DESCRIPTION (provided by applicant): The androgen receptor is the cellular mediator of the male sex hormone testosterone and a key regulator in the development and progression of prostate cancer. Androgen receptor is unusual among the family of nuclear receptors in that its hormone independent AF-1 activity is the dominant gene transactivation function. AF-1 activity has been mapped to a region of the N-terminal domain. This domain contains interaction sites for numerous coactivators, corepressors, and other proteins. Despite the critical role of AF-1 in androgen receptor function and prostate cancer pathology, the structural basis of AF-1 activity, and more generally the function of the N-terminal androgen receptor domain remains unknown. We will define, express, purify and crystallize stable complexes of N-terminal androgen receptor domain fragments with partnering domains of a representative subset of fifty coregulator proteins such as SRC's and ARA's. Our proposal employs high-throughput robotic expression and purification systems to screen thousands of paired combinations of N-terminal receptor domain and coregulator fragments to discover synergistic folding domains for stable complexes. The structure of androgen receptor-coregulator complex will reveal interactions necessary to design the first Pharmaceuticals directed to the N-terminal receptor domain in controlling prostate cancer. The ability to antagonize androgen receptor activity with agents affecting the interaction of the N-terminal androgen receptor domain with steroid receptor coactivators is expected to inhibit metastatic invasion and proliferation of prostate cancer. The goal of our study will be to establish a discovery strategy to identify small organic compounds as antagonists of androgen receptor activity in prostate cancer tumors by disrupting critical interactions between the N-terminal domain of the androgen receptor and key coregulators.