ENDOTHELIN RECEPTOR--PURIFICATION AND CHARACTERIZATION

内皮素受体——纯化和表征

• 批准号: 3087704
• 负责人: MICHAEL D GUNN
• 金额: $ 7.53万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3087704
• 关键词: Xenopus; affinity chromatography; biological signal transduction; blood vessel disorder; calcium flux; chemical structure function; cow; gel electrophoresis; heart cell; high performance liquid chromatography; hypertension; ion exchange chromatography; molecular cloning; muscle tone; nuclear magnetic resonance spectroscopy; nucleic acid probes; peptide chemical synthesis; protein purification; protein structure function; pulmonary hypertension; radionuclides; receptor binding; receptor expression; site directed mutagenesis; tissue /cell culture; vascular resistance; vascular smooth muscle; vasoconstrictors; vasospasm

Numerous cardiovascular diseases involve alterations in vascular tone and resistance to blood flow. These include systemic and pulmonary hypertension and acute coronary and cerebral artery vasospasm. It is now known that the vascular endothelium produces important vaso-regulatory substances which mediate vascular smooth muscle tone. One such substance is endothelin, the most potent vasoconstrictor polypeptide known to date. The long term goal of this proposal is to understand the role of endothelin in vascular disease and to develop methods of blocking its pathological effects. Endothelin acts by binding to high affinity receptors on the surface of vascular smooth muscle cells. The aim of this project is to purify the endothelin receptor and use it to study the mechanism of endothelin action. In order to do this, our specific aims are to determine the important structural features of the endothelin molecule, to determine the endothelin binding characteristics of various tissues, to purify, clone, and sequence the endothelin receptor, and to use mutated forms of the receptor to study endothelin's signaling pathways. This work will utilize newer biochemical techniques including the use of nuclear magnetic resonance spectroscopy to determine the three dimensional structure of endothelin and expression cloning as a means to isolate the endothelin receptor. We have the ability to synthesize functional endothelin derivatives, which may allow the production of specific endothelin antagonists. Preliminary studies have shown that endothelin receptor stimulation causes a marked increase in intracellular calcium and may be coupled to opening of the voltage activated calcium channel. Site directed mutagenesis will allow us to study the mechanism of this and other signaling pathways important to cell biology. The results of these investigations should provide a better understanding of the basic aspects of endothelin action and should aid in the development of new approaches to the study and treatment of vascular disease.

许多心血管疾病涉及血管张力和血管收缩的改变