PHARMACOLOGY OF CARDIAC A1 ADENOSINE RECEPTORS

心脏 A1 腺苷受体的药理学

• 批准号: 3348738
• 负责人: GARY L STILES
• 金额: $ 21.6万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-06 至 1995-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3348738
• 关键词: G protein; adenosine; affinity chromatography; affinity labeling; bradycardia; calcium transporting ATPase; cyclic AMP; drug receptors; glycoprotein structure; heart contraction; heart pharmacology; laboratory rat; lipolysis; molecular cloning; myocardial ischemia /hypoxia; nucleic acid sequence; protein structure; receptor coupling

The major goals of the proposed research are the elucidation of the molecular structure of the A1 adenosine receptor and the biochemical mechanism(s) involved in the transduction of information from the interaction of adenosine with the receptor to the production of intracellular signals. Adenosine is released from cells under normal and pathophysiological conditions, such as ischemia and hypoxia and can interact with A1 adenosine receptors to directly inhibit adenylate cyclase, modulate K+ channels, blunt the effect of catecholamines, induce bradycardia and AV nodal block, inhibit lipolysis and induce sedation. The mechanisms and mode of coupling of the A1 adenosine receptor (A1AR) to the various effector systems remain largely unknown. Which guanine nucleotide regulatory proteins (G proteins) the A1AR couple to and how specificity and selectivity are maintained for these diverse pathways remains to be determined. These studies will (1) complete the purification of the A1AR to homogeneity, (2) study the purified A1AR in reconstitution systems with G proteins derived from both purification of Gi and Go from bovine brain and with recombinant G protein (alpha subunits), (3) determine if the A1AR is a substrate for protein kinases and the functional consequences of phosphorylation, (4) begin to ascertain the mechanisms involved in the "tight" coupling of A1AR with G proteins and which G proteins the A1AR couples to and (5) to obtain protein sequence data from the purified A1AR to clone the A1AR using an oligonucleotide hybridization approach. These biochemical and molecular biologic approaches are complimentary to in vivo animal and cell culture receptor regulation and cell biology studies currently ongoing in the Principal Investigator's laboratory. The ultimate goal of the proposed research is to understand the structure and function of the components of the A1AR transmembrane signalling apparatus both in health and disease so that we can manipulate its components for therapeutic benefit.

拟议研究的主要目标是阐明 A1腺苷受体的分子结构及生化 涉及信息传递的机制 腺苷与受体的相互作用，产生 细胞内信号。 正常情况下细胞会释放腺苷 病理生理条件，例如缺血和缺氧，可以 与A1腺苷受体相互作用，直接抑制腺苷酸环化酶， 调节 K+ 通道，减弱儿茶酚胺的作用，诱导 心动过缓和房室结阻滞，抑制脂肪分解并诱导镇静。 这 A1 腺苷受体 (A1AR) 与 A1 腺苷受体 (A1AR) 偶联的机制和模式 各种效应器系统仍然很大程度上未知。 鸟嘌呤核苷酸是哪个 A1AR 偶联的调节蛋白（G 蛋白）以及特异性和特异性如何 对这些不同途径的选择性仍有待保持 决定。 这些研究将（1）完成A1AR的纯化 为了同质性，(2) 研究重构系统中纯化的 A1AR G 蛋白源自从牛脑中纯化的 Gi 和 Go 并使用重组 G 蛋白（α 亚基），(3) 确定 A1AR 是否 是蛋白激酶的底物，其功能后果 磷酸化，（4）开始确定参与的机制 A1AR 与 G 蛋白的“紧密”耦合以及哪些 G 蛋白与 A1AR 与 和 (5) 偶联，从纯化的 A1AR 中获取蛋白质序列数据 使用寡核苷酸杂交方法克隆 A1AR。 这些 生化和分子生物学方法与体内方法是互补的 动物和细胞培养受体调节和细胞生物学研究 目前正在首席研究员的实验室进行。 拟议研究的最终目标是了解结构 A1AR 跨膜信号传导组件的功能和功能 健康和疾病的装置，以便我们可以操纵它 具有治疗效果的成分。