P2T PURINERGIC RECEPTORS

P2T 嘌呤能受体

• 批准号: 6782593
• 负责人: JOSE L BOYER
• 金额: $ 25.46万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6782593
• 关键词: adenosine diphosphate; adenosinetriphosphatase; adenylate cyclase; cell line; enzyme activity; expression cloning; extracellular; human subject; inhibitor /antagonist; molecular cloning; nucleotidases; nucleotide metabolism; phosphorylation; platelet aggregation; protein localization; purinergic receptor; receptor coupling; receptor expression; receptor sensitivity; stoichiometry

Extracellular nucleotides interacting with specific nucleotide receptors (P2 receptors) participate in a wide variety of important physiological responses. One of the first recognized physiological roles of extracellular nucleotides was the aggregation of platelets induced by ADP. Adenine nucleotide effects of platelet function result from the activation of at least three different receptor subtypes: P2X1, P2Y1 and P2Y AC receptors. In spite of significant advances in the characterization of the receptors involved, the effects of nucleotides and associated signaling mechanisms remain poorly understood. ADP-induced platelet aggregation requires the simultaneous activation of P2Y1 and P2Y AC receptors, indicating that both receptor proteins are potential targets for antithrombotic agents. The affects of adenine nucleotides on platelet receptors are strongly influenced by the complex metabolism of extracellular nucleotides by ubiquitous ecto- nucleotidases. Therefore, an integrated study of receptor signaling and nucleotide metabolism is necessary to understand the regulation of platelet function by extracellular nucleotides. We have studied extensively in C5 rat glioma cells a P2Y receptor coupled to inhibition of adenylyl cyclase, and confirmed that the pharmacological and signaling properties of this receptor are identical to those of the platelet P2Y AC receptor. In spite of great efforts, the molecular identification of both, the platelet and C6 cell P2Y AC receptors remain elusive. The aim of this research is to clone the platelet ADP receptor coupled to inhibition of adenylyl cyclase, and to establish the physiological role of the extracellular nucleotide metabolism on P2Y receptor signaling. A research plan for the cloning of the platelet P2Y AC receptor involving expression-cloning strategies using the c6 P2Y AC receptor as model system are outlined. The study of extracellular nucleotide metabolism on the signaling of platelet and other P2Y receptors will follow from the progress of the previous funding period. We are now in the position to ask molecular questions about the functional interaction of P2Y receptors and extracellular nucleotide metabolism. Cloning the P2Y AC receptor and understanding the regulation of nucleotide metabolism are essential steps towards the understanding of the complex molecular mechanism of ADP-mediated platelet activation. This information should prove useful in the development of new biochemical and pharmacological tools with potential use in the treatment of coagulation-related diseases.

与特定核苷酸受体（P2受体）相互作用的细胞外核苷酸参与了多种重要的生理反应。 细胞外核苷酸的第一个公认的生理作用之一是ADP诱导的血小板的聚集。血小板功能的腺嘌呤核苷酸作用至少激活三种不同的受体亚型：P2X1，P2Y1和P2Y AC受体。 尽管在涉及受体的表征方面取得了重大进展，但核苷酸和相关信号传导机制的影响仍然很少了解。 ADP诱导的血小板聚集需要同时激活P2Y1和P2Y AC受体，这表明两种受体蛋白都是抗血栓形成剂的潜在靶标。 腺嘌呤核苷酸对血小板受体的影响受到无处不在的核核苷酸酶对细胞外核苷酸的复杂代谢的强烈影响。 因此，必须对受体信号传导和核苷酸代谢进行综合研究，以了解细胞外核苷酸对血小板功能的调节。 我们已经在C5大鼠神经胶质瘤细胞中进行了广泛的研究，该受体与抑制腺苷酸环化酶相结合，并证实该受体的药理和信号传导特性与血小板P2Y AC受体的药理和信号传递相同。 尽管做出了巨大的努力，但同时识别的分子鉴定，血小板和C6细胞P2Y AC受体仍然难以捉摸。 这项研究的目的是克隆血小板ADP受体结合抑制腺苷酸环化酶，并确定细胞外核苷酸代谢在P2Y受体信号传导上的生理作用。 概述了涉及使用C6 P2Y AC受体作为模型系统的涉及表达克隆策略的血小板P2Y AC受体克隆的研究计划。 对血小板和其他P2Y受体信号传导的细胞外核苷酸代谢的研究将取决于上一流的资金时期的进展。 现在，我们可以询问有关P2Y受体和细胞外核苷酸代谢的功能相互作用的分子问题。 克隆P2Y AC受体并了解核苷酸代谢的调节是了解ADP介导的血小板激活的复杂分子机制的重要步骤。 该信息应证明可用于开发新的生化和药理学工具，并可能在治疗凝血相关疾病方面使用。