REGULATION OF KININ DELIVERY ON HUVEC

HUVEC 上激肽传递的调节

• 批准号: 2430754
• 负责人: ALVIN H SCHMAIER
• 金额: $ 28.32万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-15 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2430754
• 关键词: bradykinin; human tissue; kallikreins; keratin; kininogens; kinins; membrane proteins; molecular cloning; protein purification; protein sequence; protein transport; receptor binding; receptor expression; site directed mutagenesis; tissue /cell culture; vascular endothelium; vasodilation; zymogens

The hypothesis that forms the basis of this proposal is that the endothelial cell receptor for the plasma kininogens is a multimeric structure that orders contact protein zymogen assembly and activation of prokallikreins for sequestered kinin liberation and other kallikrein- dependent biologic activities. Expression of kininogens, both high (HK) and low (LK) molecular mass kininogens on this receptor permits kinins [bradykinin (BK) and met-lys-bradykinin (kallidin)] to be liberated to influence local vascular biology. This hypothesis is based upon the finding that endothelial cells contain several membrane-expressed kininogen binding proteins. Second, both HK and LK themselves have multiple binding-domains for its particular orientation onto its endothelial cell binding site(s), putative receptor(s). Third, HK serves as the endothelial cell binding site for prekallikrein. Fourth, prekallikrein only bound to HK on endothelium is activated by an endothelial cell, calcium requiring metalloprotease for kinetically favorable pro-urokinase and subsequent plasminogen activation. To test the above hypothesis, the specific aims of this proposal are as follows: (1) The kininogen receptor(s) on endothelial cells will be determined. (2) The mechanism(s) regulating kinin liberation from cell- bound kininogens will be determined. (3) The fine mapping of sequences on kininogens that participate in its binding to endothelium will be characterized. These investigations will characterize how kinins are liberated from its parent proteins in the intravascular compartment to influence vascular biology. These studies will provide a new avenue for modulation of all bradykinin-dependent, endothelial cell activities such as tPA liberation, NO formation, elevation of cGMP, prostacyclin synthesis, superoxide formation, and smooth muscle hyperpolarization factor production. Modulation of bradykinin liberation is important in the regulation of local blood pressure in coronary, renal, pulmonary, and cerebral circulations.

构成本提案基础的假设是 血浆激肽原的内皮细胞受体是一种多聚体 命令接触蛋白酶原组装和激活的结构 用于释放隔离的激肽和其他激肽释放酶的前激肽释放酶- 依赖的生物活性。激肽原的表达，均高（HK） 该受体上的低（LK）分子量激肽原允许激肽 [缓激肽 (BK) 和 met-lys-缓激肽 (激肽)] 被释放到 影响局部血管生物学。 该假设基于以下发现：内皮细胞含有 几种膜表达的激肽原结合蛋白。二、既是香港 并且 LK 本身具有多个结合域，用于其特定的 假定的内皮细胞结合位点的方向 受体。第三，HK作为内皮细胞结合位点 前激肽释放酶。第四，前激肽释放酶仅与内皮细胞上的 HK 结合 由内皮细胞激活，钙需要金属蛋白酶 动力学有利的尿激酶原和随后的纤溶酶原 激活。 为了检验上述假设，本提案的具体目标是 如下： (1) 内皮细胞上的激肽原受体将是 决定。 (2) 调节激肽从细胞中释放的机制 将测定结合的激肽原。 (3) 序列的精细映射 参与其与内皮细胞结合的激肽原将是 特点。 这些研究将描述激肽如何从其 血管内室中的母蛋白影响血管 生物学。这些研究将为所有的调节提供新途径 缓激肽依赖性内皮细胞活性，例如 tPA 释放， NO 形成、cGMP 升高、前列环素合成、超氧化物 形成和平滑肌超极化因子的产生。 缓激肽释放的调节对于调节 冠状动脉、肾动脉、肺动脉和脑动脉的局部血压 流通量。