INHIBITION OF GRIP1 & HTR INTERACTIONS

我们这个项目的总体目标是建立方法， 阻断蛋白质与小有机分子的相互作用。 这种抑制剂可以选择性地阻止特定的 复合物，从而破坏下游过程。 往往如此复杂 形成是由模块化蛋白结合模块介导的， WW、PDZ和SH 3基序。 最近，一些结构研究 揭示了一类新的蛋白质相互作用。 这种绑定模式 包括一个小螺旋段的一个面的装配， 蛋白质进入另一个明确的裂缝。 一个例子是 糖皮质激素受体相互作用蛋白1（GRIP 1）与 人甲状腺激素受体β 1（hTR= DF 1）配体（T3） 复杂. 由于相对较小的总结合位点， 螺旋和两者之间没有主链氢键 合作伙伴，这种类型的互动是一个有前途的测试案例 用于蛋白质-蛋白质结合的小分子抑制剂。 长 该项目的长期研究目标是开发一种细胞渗透性 GRIP 1/hTRb 1/T3结合的抑制剂，特别是（1）开发一种 适用于高通量筛选的稳健的生物化学测定 GRIP 1与hTR= DF 1/T-3复合物结合的竞争性抑制剂; (2)合成构象受限的α-螺旋肽 跨越GRIP 1的假定结合部分，并测试此 模板作为抑制剂;（3）探索最小结构 GRIP 1螺旋与hTR结合的限制条件= DF 1/T-3 （4）开始定义结构-活性 使用肽模型的GRIP 1螺旋的关系。 计算机 图形实验室的资源被用来分析 配体与受体相互作用和设计电势 基于这种分析的抑制剂。

Our overall goal of this project is to establish methods for blocking protein-protein interactions with small organic molecules. Such inhibitors could selectively prevent the assembly of particular complexes and thus disrupt downstream processes. Often such complex formations are mediated by modular protein binding modules such as the WW, PDZ, and SH3 motifs. Recently, several structural studies have revealed a new class of protein interaction. This binding mode involves the fitting of one face of a small helical segment of one protein into a well-defined cleft on the other. One example is the binding of glucocorticoid receptor-interacting protein 1 (GRIP1) to the human thyroid hormone receptor beta 1 (hTR=DF1) ligand (T3) complex. Due to the rather small size of the overall binding site for the helix and the absence of backbone hydrogen bonding between the two partners, this type of interaction represents a promising test case for small molecule inhibitors of protein-protein binding. The long term research goal of the project is to develop a cell permeable inhibitor for GRIP1/hTRb1/T3 binding, specifically (1) to develop a robust biochemical assay suitable for high throughput screening for competitive inhibitors of the binding of GRIP1 to hTR=DF1/T-3 complex; (2) to synthesize a conformationally constrained a-helical peptide that spans the putative binding portion of GRIP1 and test this template as an inhibitor; (3) to explore the minimal structural constraints for the binding of the GRIP1 helix to the hTR=DF1/T-3 complex; and, (4) to begin to define the structure-activity relationship for the GRIP1 helix using peptide models. The Computer Graphics Laboratory resources are being utilized to analyze the interaction between ligand and receptor and to design potential inhibitors based upon this analysis.