GPCR Network

描述(由申请人提供)：G蛋白偶联受体可感知各种惊人的细胞外分子信号，并触发复杂的细胞内和生理反应。它们共享一个由七个跨膜螺旋组成的共同结构，这些螺旋由广泛的细胞内和细胞外环和终端域连接。最近首次对人肾上腺素能受体、火鸡能肾上腺素能受体和人腺苷A2a受体进行了高分辨率研究，证明了该蛋白家族结构确定的可行性。建立了膜蛋白结构确定中心(CMPD)，以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个具有代表性的GPCR的高分辨率结构。小分子配体受体、肽和蛋白质受体、脂类受体、B-F类受体以及处于活动和非活动功能状态的受体都需要具有生物相关粒度的受体结构。将用一组不同的药理配基确定每个受体的结构，以确定受体结合部位(S)。将用纯化的受体与不同的配体结合进行溶液研究，以了解受体的动力学，使用氢-氚交换和核磁共振波谱。与NIH筛选中心合作，将使用一个小分子探针库来分析每个受体，并使用高通量热稳定性筛选发现变构结合位点。通过生物知情的代表性受体的选择，我们将通过外部合作者对接近的同源物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。生成的数据将在与蛋白质结构倡议指导方针一致的时间框架内提供给社区。将通过现场培训、讲习班、会议和出版物实现技术获取。通过为PSI：生物网络提名目标提供30%的管道能力承诺，将提供对CMPD核心设施的处理访问。根据CMPD研究人员的经验，优先选择人类或真核膜蛋白来最大限度地利用CMPD能力。 公共卫生相关性：G蛋白偶联受体是人类基因组中最大的蛋白质家族，有800多个不同的成员，超过50%的治疗药物是靶标。细胞表面受体的遗传变异会导致与多种人类疾病有关的功能中断。为了了解这个重要的蛋白质家族的分子识别和信号转导特性，对于基础和应用生物医学研究来说，结构研究是必要的。

DESCRIPTION (provided by applicant): 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 ¿2 adrenergic, turkey ¿1 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. PUBLIC HEALTH RELEVANCE: G-protein coupled receptors are the largest protein family in the human genome with more than 800 different members and the target for more than 50% of all therapeutic drugs. Genetic variation in the cell surface receptors result in the disruption of functions that are implicated in a wide variety of human diseases. Structural studies are necessary to understand the exquisite molecular recognition and signal transduction properties of this important protein family for both basic and applied biomedical research.