GPCR Network

G蛋白偶联受体能感知多种细胞外分子信号， 复杂的细胞内和生理反应。它们有一个共同的结构， 跨膜螺旋通过广泛的细胞内和细胞外环连接， 域.该蛋白质家族的结构确定的可行性最近被证明与第一个 人2肾上腺素能、火鸡1肾上腺素能和人腺苷A2 A的高分辨率研究 受体。膜蛋白结构测定中心（CMPD）已经建立， 蛋白质家族特异性平台，以确定15-20个代表性GPCR的高分辨率结构 分布在整个系统发育树中。受体结构需要在生物学相关的 粒度，用于小分子配体受体、肽和蛋白质受体、脂质受体，B-F类 受体，以及处于活性和非活性功能状态的受体。每个受体结构将 用一组不同的药理学配体测定以限定受体结合位点。溶液 将用与不同配体结合的纯化受体进行研究，以了解受体 动力学使用氢-氘交换和NMR光谱。与NIH合作 筛选中心，一个小分子探针库将用于分析每个受体，并发现 使用高通量热稳定性筛选来确定变构结合位点。通过一个生物学上知情的 选择有代表性的受体，我们将最大限度地提高CMPD的影响，通过计算 通过外部合作者对密切同源物和功能研究进行建模，从而建立PSI GPCR Network.生成的数据将在符合 蛋白质结构倡议组织（Protein Structure Initiative）将通过现场培训获得技术， 讲习班、会议和出版物。将提供对CMPD核心设施的处理访问 通过30%的管道能力承诺的PSI：生物网络提名的目标。基于 根据CMPD研究者的经验，优选人或真核细胞膜蛋白， 最大限度地利用CMPD功能。

G protein-coupled receptors sense an astonishing variety of extracellular molecular signals and trigger complex intracellular and physiological responses. They share a common architecture of seven transmembrane helices connected by a broad range of intra- and extra-cellular loops and terminal domains. Structure determination feasibility of this protein family was demonstrated recently with the first high resolution studies on the human β2 adrenergic, turkey β1 adrenergic, and human adenosine A2A receptors. The Center for Membrane Protein Structure Determination (CMPD) has been created to use a protein family specific platform to determine the high resolution structures of 15-20 representative GPCRs distributed across the phylogenetic tree. Receptor structures are needed at a biologically relevant granularity, for small molecule ligand receptors, peptide and protein receptors, lipid receptors, class B-F receptors, and of receptors in the active and inactive functional states. Each receptor structure will be determined with a set of different pharmacological ligands to define the receptor binding site(s). Solution studies will be conducted with purified receptors bound to different ligands to understand receptor dynamics using hydrogen-deuterium exchange and NMR spectroscopy. In collaboration with the NIH screening center, a library of small molecule probes will be used to analyze each receptor and discover allosteric binding sites using a high throughput thermal stability screen. Through a biologically informed selection of representative receptors, we will maximize the CMPD‟s impact through computational modeling of close homologs and functional studies by external collaborators thereby establishing The PSI GPCR Network. The generated data will be provided to the community in a time frame consistent with the guidelines of the Protein Structure Initiative. Technology access will be achieved through on-site training, workshops, meetings, and publications. Processing access to the CMPD core facility will be provided through a 30% pipeline capacity commitment for the PSI:Biology Network nominated targets. Based on the experience of the CMPD investigators, preference will be for human or eukaryotic membrane proteins to maximally leverage the CMPD capabilities.