Van Yüzüncü Yıl University, School of Health, Department of Nutrition and Dietetics, Van, Turkey.
Van YüzüncüYıl University, Faculty of Science, Department of Chemistry, Van, Turkey.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jan 1;1104:190-195. doi: 10.1016/j.jchromb.2018.11.023. Epub 2018 Nov 19.
Angiotensin converting enzyme (ACE, peptidyldipeptidase A, EC 3.4.15.1) plays an important role in the regulation of blood pressure. In this study, ACE was purified from human plasma by affinity chromatography in single step. The enzyme purified in 5367-fold from human plasma and specific activity was found to be 1208 EU/mg protein. The purity and molecular weight of ACE were determined by SDS-PAGE, which indicated two bands at around 60 kDa and 70 kDa on the gel. Effect of oxidized glutathione (GSSG) peptide and reduced glutathione (GSH) peptide on purified ACE activity were also investigated in which lisinopril was used as reference inhibitor. GSSG showed activation effect on ACE activity whereas GSH provided inhibition effect. In the lights of activity (%) versus activator graph for GSSG and activity (%) versus inhibitor graphs for GSH and lisinopril; IC values for GSH and lisinopril were determined to be 16.2 μM and 0.781 nM, respectively. Type of inhibition for GSH and lisinopril from graph Lineweaver-Burk was found to be reversible non-competitive inhibition and K constants for GSH and lisinopril were calculated as 11.7 μM and 0.662 nM, respectively.
血管紧张素转换酶（ACE，肽二肽酶 A，EC 3.4.15.1）在血压调节中起着重要作用。在这项研究中，ACE 从人血浆中通过亲和层析在一步中进行了纯化。该酶从人血浆中纯化了 5367 倍，比活为 1208 EU/mg 蛋白。通过 SDS-PAGE 确定 ACE 的纯度和分子量，凝胶上大约 60 kDa 和 70 kDa 处有两条带。还研究了氧化型谷胱甘肽（GSSG）肽和还原型谷胱甘肽（GSH）肽对纯化 ACE 活性的影响，其中赖诺普利用作参考抑制剂。GSSG 对 ACE 活性表现出激活作用，而 GSH 则提供抑制作用。根据 GSSG 的活性（%）与激活剂图和 GSH 及赖诺普利的活性（%）与抑制剂图；确定 GSH 和赖诺普利的 IC 值分别为 16.2 μM 和 0.781 nM。从图 Lineweaver-Burk 来看，GSH 和赖诺普利的抑制类型为可逆非竞争性抑制，GSH 和赖诺普利的 K 常数分别计算为 11.7 μM 和 0.662 nM。
