PEPTIDE PROBES OF THE 5HT1A RECEPTOR/G PROTEIN INTERFAC

描述（改编自申请人摘要）： 5-羟色胺（5-HT）受体参与人类健康的关键方面， 抑郁症和偏头痛等疾病。我们的长期目标是研究 这些受体的分子药理学，特别是 5-羟色胺受体和一个重要的细胞信号转导方案，G 蛋白质系统在本项目中，我们提出了一种人类的研究， 5-羟色胺受体，称为5 HTla受体。这种受体属于一种 包括其它重要模型受体的受体大超家族 如肾上腺素能、多巴胺能和毒蕈碱受体。基于已知 这些模型受体和5-羟色胺受体本身的特性，我们有 合成并表征了一种来自假定G蛋白偶联的肽， 5 HTla受体的区域。在本研究中，我们将利用固相 合成这种生物活性物质的结构变体的方法 肽。将使用激动剂抑制测定来测量变体的活性。 进一步的功能测定将包括测定第二个细胞的变化。 信使环AMP或腺苷酸环化酶，并通过测量 鸟苷三磷酸（GTP）/ G蛋白结合。GTP结合试验将 在粗细胞制剂以及克隆的G蛋白中进行 系统圆二色性（CD）研究将分析溶液结构， 优先肽。结合和功能实验的信息将 有助于确定关键分子的最佳肽工具 接口的药理学研究。先进之处在于分析和建模 分子界面创造了一个现实的机会， 受体/G蛋白接触表面的构象。如此选择的肽 将作为G蛋白的探针，利用多维核磁共振技术， 核磁共振（NMR），以更好地了解受体-G的活性构象 蛋白质界面从我们的研究信息将增加必要的知识， 模拟受体超家族。开发新的治疗药物可能 跟随.

DESCRIPTION (Adapted from applicant's abstract): Serotonin (5HT) receptors are involved in critical aspects of human health and disease such as depression and migraine. Our long-term goals are to study the molecular pharmacology of these receptors, especially interactions between serotonin receptors and a significant cellular signal transducing scheme, the G protein system. In the present project, we propose studies with a type of human serotonin receptor known as the 5HTla receptor. This receptor belongs to a large superfamily of receptors which include other important model receptors such as adrenergic, dopaminergic, and muscarinic receptors. Based upon known properties of these model receptors and serotonin receptors themselves, we have synthesized and characterized a peptide from a putative G protein coupling region of the 5HTla receptor. In the present study we will utilize solid phase methods for synthesizing structural variants of this biologically active peptide. Activity of variants will be measured using agonist inhibition assays. Further functional assays will include determinations in changes of the second messenger cyclic AMP or the enzyme adenylyl cyclase, and by measurement of guanosine triphosphate (GTP)/ G protein binding. GTP binding assays will be performed in crude cellular preparations as well as in a cloned G protein system. Circular dichroism (CD) studies will analyze solution structures of priority peptides. Information from binding and functional experiments will contribute to determination of the best peptide tools for key molecular pharmacology studies of the interface. Advancements is analyzing and modeling molecular interfaces create a realistic opportunity for discovery of active conformations of receptor/G protein contact surfaces. Thus selected peptides will be used as probes of G protein using multi-dimensional nuclear magnetic resonance (NMR) to better understand the active conformations of the receptor-G protein interface. Information from our studies will add essential knowledge to modeling the receptor superfamily. Development of novel therapeutic drugs may follow.