Folding of Androgen Receptor-Coregulator Complex

描述(申请人提供)：雄激素受体是男性性激素睾酮的细胞介体，也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寻常的，因为它的激素非依赖性的AF-1活性是主导的基因反式激活功能。AF-1的活性已被映射到N-末端结构域的一个区域。这个结构域包含许多辅活化子、辅阻遏子和其他蛋白质的相互作用部位。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，更广泛地说，N端雄激素受体结构域的功能仍不清楚。我们将定义、表达、纯化和结晶稳定的N-末端雄激素受体结构域片段与50个具有代表性的辅调节蛋白子集的合作结构域，例如SRC‘s和ARA’s。我们的建议利用高通量机器人表达和纯化系统来筛选数千对N-末端受体结构域和辅助调节蛋白片段的配对组合，以发现稳定复合体的协同折叠结构域。雄激素受体-辅助调节复合体的结构将揭示设计针对N端受体结构域控制前列腺癌的第一种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剂相互作用的药物来拮抗雄激素受体活性的能力有望抑制前列腺癌的转移侵袭和增殖。我们研究的目标将是建立一种发现策略，通过破坏雄激素受体的N末端结构域和关键辅助调节因子之间的关键相互作用，确定小的有机化合物作为前列腺癌肿瘤中雄激素受体活性的拮抗剂。

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.