INHIBITION OF GRIP1 & HTR INTERACTIONS

我们这个项目的总体目标是建立 阻止蛋白质与有机小分子的相互作用。 这种抑制剂可以选择性地阻止特定的 并因此扰乱下游进程。往往如此复杂 形成由模块化的蛋白质结合模块介导，如 WW、PDZ和SH3图案。最近，几项结构研究已经 揭示了一类新的蛋白质相互作用。此绑定模式 包括对一个小螺旋线段的一个面进行拟合 蛋白质在另一端形成一个清晰的裂隙。一个例子是 糖皮质激素受体相互作用蛋白1与糖皮质激素受体相互作用蛋白1的结合 人甲状腺激素受体β1(HTR=DF1)配体(T3) 很复杂。由于总结合部位的大小相当小，因此 螺旋和主链之间没有氢键 合作伙伴，这种类型的交互代表了一个很有前途的测试用例 用于蛋白质-蛋白质结合的小分子抑制剂。《长河》 该项目的学期研究目标是开发一种可渗透的细胞 GRIP1/hTRb1/T3结合的抑制剂，特别是(1)开发一种 适用于高通量筛选的强健生化方法 GRIP1与HTR=DF1/T-3复合体结合的竞争性抑制剂； (2)合成构象受限的α-螺旋肽 它跨越了GRIP1的假定结合部分，并测试了这一点 模板作为抑制剂；(3)探索最小结构 GRIP1螺旋与HTR=DF1/T-3结合的限制条件 复杂性；以及，(4)开始定义结构-活动 用多肽模型研究GRIP1螺旋之间的关系。这台电脑 正在利用图形实验室的资源来分析 配体与受体的相互作用及其设计电势 基于这一分析的抑制剂。

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.