GPCR Network

G蛋白偶联受体感知细胞外分子信号的惊人变化并触发 复杂的细胞内和生理反应。它们共享一个由七个人组成的公共架构 跨膜螺旋由广泛的细胞内环和细胞外环和终末连接 域名。这个蛋白质家族的结构确定的可行性最近被证明是第一次 人肾上腺素能、火鸡1肾上腺素能和人腺苷A2a的高分辨率研究 感受器。膜蛋白结构测定中心(CMPD)已经成立，以使用 蛋白质家族特异性平台用于确定15-20个具有代表性的GPCR的高分辨结构 分布在系统发育树上。在生物相关的情况下需要受体结构 B-F类小分子配体受体、肽和蛋白质受体、脂类受体的粒度 受体，以及处于活动和非活动功能状态的受体。每个受体结构将是 用一组不同的药理配基确定受体的结合部位(S)。解 将用纯化的受体结合不同的配体进行研究，以了解受体 使用氢-氚交换和核磁共振波谱的动力学。与美国国立卫生研究院合作 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的变构结合位点。通过生物学上了解到的 选择有代表性的受体，我们将通过计算最大限度地影响CMPD&S 外部合作者对密切的同源基因和功能研究进行建模，从而建立PSI GPCR网络。生成的数据将在符合以下规定的时间内提供给社区 蛋白质结构倡议指南。将通过现场培训实现技术准入， 研讨会、会议和出版物。将提供对CMPD核心设施的处理通道 通过30%的管道能力承诺为PSI：生物网络提名目标。基于 根据CMPD研究人员的经验，将优先选择人类或真核膜蛋白 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.