THE INHIBITION OF GRIP1/HTR INTERACTIONS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们这个项目的总体目标是建立阻断蛋白质与小有机分子相互作用的方法。这种抑制剂可以选择性地阻止特定复合物的组装，从而破坏下游过程。通常，这种复合物的形成是由模块化蛋白结合模块如WW、PDZ和SH 3基序介导的。最近，一些结构研究揭示了一类新的蛋白质相互作用。这种结合模式涉及一种蛋白质的小螺旋片段的一个面与另一个蛋白质的明确裂缝的配合。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）复合物的结合。由于螺旋的整体结合位点的尺寸相当小，并且两个伴侣之间不存在骨架氢键，这种类型的相互作用代表了蛋白质-蛋白质结合的小分子抑制剂的有希望的测试案例。本项目的长期研究目标是开发GRIP 1/hTRb 1/T3结合的细胞渗透性抑制剂，具体而言，（1）开发适用于高通量筛选GRIP 1与hTR= DF 1/T-3复合物结合的竞争性抑制剂的稳健的生物化学测定法;（2）合成跨越GRIP 1的推定结合部分的构象受限的α-螺旋肽，并测试该模板作为抑制剂;（3）探索GRIP 1螺旋与hTR= DF 1/T-3复合物结合的最小结构限制;（4）开始使用肽模型定义GRIP 1螺旋的结构-活性关系。计算机图形实验室的资源正在被用来分析配体和受体之间的相互作用，并设计基于此分析的潜在抑制剂。

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. Our overall goal for 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's resources are being utilized to analyze the interaction between ligand and receptor and to design potential inhibitors based upon this analysis.