KININ RECEPTORS--MOLECULAR PROPERTIES AND REGULATION

这笔拨款是为了支持一项基础研究计划而申请的 旨在从分子水平上阐明病毒的性质、模式 受体的功能和调节的分子机制 对于激动素。主要的哺乳动物激动素，缓激肽和卡利丁， 从大分子量的血浆和组织中产生 激动素，激肽原，通过特定的丝氨酸蛋白酶，激肽释放酶， 例如对组织损伤和炎症反应作出反应。在……里面 活体，激肽作用的主要结果是 小动脉血管扩张引起的血压，小动脉扩张引起的水肿 血管通透性增加，疼痛。的确，激肽可以模仿 所有炎症的主要征兆。此外，激动素还具有 与心血管休克的发生有关，高... 紧张和类风湿性关节炎。在细胞水平上，激动素 刺激肌醇磷脂代谢，然后增加 细胞内钙水平，花生四烯酸和花生四烯酸的释放 前列腺素通过刺激磷脂酶C和 A2。这项研究计划有四个主要目标 旨在加深对激动素功能的了解 多种生理和病理生理学中的受体 条件。目标是：1)发展高亲和力 激动素受体的放射性配体；2)阐明分子 激动剂占用的翻译机制 细胞外部的受体进入到产生 细胞内信号。最初，我们会继续进行这些研究。 使用培养的血管平滑肌细胞系DDT1 MF-2。这个 激动素受体结合特性及受体激活 磷脂酶A2和C的研究将在膜和 完整的细胞制备；3)阐明分子性质 激动素受体蛋白。这一目标涉及发展 工具，如固体受体亲和树脂和亲和力 光亲和探针。然后，这些工具将用于 受体的纯化及鉴定和鉴定 受体配基结合肽的特性；4)至 阐明参与调控的分子机制 激动素受体功能。这一目标包括确定 各种因素，如同源和异源激动剂， 促进肿瘤的佛波酯、有害刺激和代谢 可能调节激动素受体反应性的状态 这种监管背后的机制。本报告中提出的目标 建议构成以下长期目标不可分割的一部分 这个实验室是为了了解分子机制 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.