THE ANGIOTENSIN CONVERTING ENZYME

血管紧张素转换酶

• 批准号: 3336779
• 负责人: IRWIN B WILSON
• 金额: $ 11.55万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-04-01 至 1987-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3336779
• 关键词: affinity chromatography; amniotic fluid; brain; cerebrospinal fluid; chemical synthesis; cyanides; enzyme structure; enzyme substrate; gel electrophoresis; high performance liquid chromatography; human tissue; lung; metalloproteins; molecular site; peptidyl dipeptidase A; pyrophosphates; spectrometry

We intend to continue our studies on the molecular, structural, kinetic, and mechanistic properties of the angiotensinI-converting enzyme. The specific aim of this grant are: 1) To obtain the limit peptide of the converting enzyme that contains some or all of the active-site residues and to sequence this peptide. 2) To study the chemical mechanism of the converting enzyme to establish whether an acyl-enzyme intermediate is formed. 3) To study the nature of the active site bindingr pocket subsites by the kinetic study of substrates and inhibitors; to study the phenomenon of substrate inhibition, the requirement for monovalent anions, and the significance of inhibition by cyanide and pyrophosphate. 4) To develop a substrate that can be used for an activity stain to detect very low levels of converting enzyme activity. 5) To continue to develop a biospecific affinity ligand for the converting enzyme and to continue to improve the general method of affinity chromatography purification. 6) To establish the existence of converting enzyme in brain tissue and other tissue sources such as cerebrospinal fluid and aminotic fluid. 7) To study the molecular and kinetic properties of the converting enzyme purified from different sources.

我们打算继续我们的分子，结构，动力学， 以及血管紧张素转换酶的机械特性。这个 这笔赠款的具体目的是： 1)获得含有一定量的转化酶的限制肽 或所有活性部位残基，并对该多肽进行测序。 2)研究转化酶的化学作用机理，建立 是否形成了酰基酶中间体。 3)研究活性中心结合的口袋亚基的性质 底物和缓蚀剂的动力学研究；研究 底物抑制，对单价阴离子的要求，以及 氰化物和焦磷酸盐抑制的意义。 4)开发一种可用于活性染色检测的底物 转化酶活性很低。 5)继续开发用于转化的生物特异性亲和配体 酶和亲和力继续提高的一般方法 层析纯化。 6)证实脑组织中存在转换酶，并 其他组织来源，如脑脊液和羊水。 7)研究转化酶的分子和动力学性质 从不同的来源提纯。