KININ RECEPTORS--MOLECULAR PROPERTIES AND REGULATION

这笔赠款是为了支持一项基础研究计划 目的是在分子水平上阐明 以及受体调节的分子机制 对于激肽。 主要的哺乳动物激肽，缓激肽和胰激肽， 在血浆和组织中由大分子量 激素，激肽原，通过特定的丝氨酸蛋白酶，激肽释放酶， 响应于例如组织损伤和炎症反应。 在 在体内，激肽作用的主要结果是， 血压由于小动脉血管扩张，水肿由于 增加血管渗透性和疼痛。 实际上，激肽模仿 所有炎症的主要症状 此外，激肽具有 与心血管休克的发生有关， 紧张和类风湿性关节炎。 在细胞水平，激肽 刺激肌醇磷脂代谢，随后增加 细胞内Ca2+水平和花生四烯酸的释放， 通过刺激磷脂酶C的活性， A2，分别。 该研究计划有四个主要目标， 目的是增加对激肽功能的了解 受体在各种生理和病理生理 条件 目标是：1）培养高亲和力 激肽受体的放射性配体; 2）阐明分子 参与翻译激动剂占据的机制 细胞外部的受体转化为 细胞内信号 在初期，这些研究将继续进行， 使用培养的平滑肌细胞系DDT1 MF-2。 的 激肽受体结合和受体激活的特性 将研究磷脂酶A2和C在膜和 完整细胞制备; 3）阐明分子特性 激肽受体蛋白。 这一目标涉及发展 工具如固体受体亲和树脂和亲和和 光亲和探针 这些工具将用于 受体的纯化和鉴定， 受体配体结合肽的表征; 4） 阐明参与调节的分子机制， 激肽受体功能 这一目标包括确定 各种因子如同源和异源激动剂， 促进肿瘤的佛波醇酯、有害刺激和代谢 可能调节激肽受体反应性的状态， 这种监管的基础机制。 本报告中提出的目标 建议是长期目标的组成部分， 这个实验室是为了了解 血管活性激素的作用。

This grant is requested to support a program in basic research aimed at elucidating at a molecular level the nature, mode of functioning, and molecular mechanisms of regulation of receptors for kinins. The major mammalian kinins, bradykinin and kallidin, are generated in plasma and tissues from large molecular weight hormogens, kininogens, by specific serine proteases, kallikreins, in response to e.g. tissue damage and inflammatory reactions. In vivo, the predominant result of kinin action is a dramatic fall in blood pressure due to arteriolar vasodilatation, edema due to increased vascular permeability, and pain. Indeed, kinins mimic all the cardinal signs of inflammation. In addition, kinins have been implicated in the genesis of cardiovascular shock, hyper- tension, and rheumatoid arthritis. At the cellular level, kinins stimulate inositol phospholipid metabolism followed by increased intracellular Ca2+ levels, and the release of arachidonic acid and prostaglandins by stimulating the activity of phospholipase C and A2, respectively. This research proposal has four major goals all aimed at an increased understanding of the function of kinin receptors in various physiological and pathophysiological conditions. The goals are: 1) to develop high affinity radioligands for kinin receptors; 2) to elucidate the molecular mechanisms involved in translation of agonist occupancy of receptors on the exterior of the cell into generation of intracellular signals. Initially, these studies will be pursued using the cultured smooth muscle cell line, DDT1 MF-2. The characteristics of kinin receptor binding and receptor activation of phospholipase A2 and C will be studied both in membrane and intact cell preparations; 3) to elucidate the molecular properties of the kinin receptor protein. This goal involves development of tools such as solid receptor affinity resins and affinity and photoaffinity probes. These tools will then be used for purification of the receptor and identification and characterization of the receptor ligand binding peptide; 4) to elucidate the molecular mechanisms involved in the regulation of kinin receptor function. This goal involves identification of various factors such as homologous and heterologous agonists, tumor-promoting phorbol esters, noxious stimuli, and metabolic state that may regulate the kinin receptor responsiveness and the mechanisms underlying such regulation. The goals presented in this proposal constitute an integral part of the long term objective of this laboratory which is to understand the molecular mechanisms of action of vasoactive hormones.