GPCR Network

G蛋白偶联受体感知细胞外分子信号的惊人多样性，并触发复杂的细胞内和生理反应。它们共有7个跨膜螺旋结构，由广泛的细胞内和细胞外环和末端结构域连接。该蛋白家族结构测定的可行性最近首次在人类β 2肾上腺素能受体、火鸡β 1肾上腺素能受体和人类腺苷A2A受体上进行了高分辨率研究。膜蛋白结构测定中心（CMPD）的建立是为了使用蛋白质家族特异性平台来确定分布在系统发育树上的15-20个代表性gpcr的高分辨率结构。对于小分子配体受体，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，需要在生物学相关的粒度上进行受体结构。每个受体结构将由一组不同的药理学配体来确定受体结合位点。溶液研究将进行纯化受体结合到不同的配体，以了解受体动力学使用氢-氘交换和核磁共振波谱。与NIH筛选中心合作，一个小分子探针库将用于分析每个受体，并使用高通量热稳定性筛选发现变弹性结合位点。通过对代表性受体的生物学选择，我们将通过外部合作者的密切同源和功能研究的计算建模来最大化CMPD的影响，从而建立PSI GPCR网络。生成的数据将在符合蛋白质结构倡议指南的时间框架内提供给社区。技术获取将通过现场培训、讲习班、会议和出版物来实现。CMPD核心设施的处理访问将通过PSI：生物网络指定目标的30%管道容量承诺提供。根据CMPD研究人员的经验，为了最大限度地利用CMPD的能力，将优先选择人或真核膜蛋白。

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 Beta2 adrenergic, turkey Beta1 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 homolog's 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.