Van YüzüncüYıl University, Faculty of Science, Department of Chemistry, Van, Turkey.
Van YüzüncüYıl University, Faculty of Science, Department of Chemistry, Van, Turkey.
Chem Biol Interact. 2021 Sep 25;347:109604. doi: 10.1016/j.cbi.2021.109604. Epub 2021 Aug 2.
Angiotensin-converting enzyme (ACE, EC 3.4.15.1) synthesized by endothelial cells and responsible for the regulation of blood pressure was purified from the bovine lung with affinity chromatography method. The purification rate of the ACE of the bovine lung was calculated as 1748- fold. Optimum pH and optimum temperature for the purified ACE were found to be 7.6 and 35-40 °C, respectively. The purity and molecular weight of the ACE were designated with SDS-PAGE. The ACE was found to have three subunits with molecular weights of 57 kDa, 66 kDa, and 190 kDa. Then, the total molecular weight of the ACE was designated as 303 kDa with gel filtration chromatography. The effects of ACE inhibitors captopril, fosinopril, lisinopril, and beta-blockers propranolol, atenolol, and diuretic triamterene on ACE activity were studied. ACE inhibitors lisinopril, captopril, fosinopril, and diuretic triamterene demonstrated an inhibition effect on ACE activity. Beta-blockers indicated no effect on ACE. IC values of captopril, fosinopril, lisinopril, and triamterene from the graphical equation were calculated as 0.835 nM, 1.159 μM, 4.085 nM, and 227 μM, respectively. The inhibition type and K values of these compounds were determined from Lineweaver-Burk plots. Captopril, fosinopril, lisinopril, and triamterene demonstrated a non-competitive inhibition effect on ACE activity. K constants were found as 1.057 nM, 1.675 μM, 6.449 nM, and 419.5 μM, respectively. Captopril indicated the highest inhibitor effect with an IC value of 0.835 nM.
血管紧张素转换酶（ACE，EC 3.4.15.1）由内皮细胞合成，负责调节血压，用亲和层析法从牛肺中纯化。牛肺 ACE 的纯化率计算为 1748 倍。发现纯化 ACE 的最适 pH 和最适温度分别为 7.6 和 35-40°C。SDS-PAGE 确定 ACE 的纯度和分子量。ACE 被发现有三个亚基，分子量分别为 57 kDa、66 kDa 和 190 kDa。然后，用凝胶过滤层析法将 ACE 的总分子量指定为 303 kDa。研究了 ACE 抑制剂卡托普利、福辛普利、赖诺普利和β-受体阻滞剂普萘洛尔、阿替洛尔和利尿剂三氨蝶呤对 ACE 活性的影响。ACE 抑制剂赖诺普利、卡托普利、福辛普利和利尿剂三氨蝶呤对 ACE 活性表现出抑制作用。β-受体阻滞剂对 ACE 无影响。从图形方程中计算出卡托普利、福辛普利、赖诺普利和三氨蝶呤的 IC 值分别为 0.835 nM、1.159 μM、4.085 nM 和 227 μM。从 Lineweaver-Burk 图确定这些化合物的抑制类型和 K 值。卡托普利、福辛普利、赖诺普利和三氨蝶呤对 ACE 活性表现出非竞争性抑制作用。K 常数分别为 1.057 nM、1.675 μM、6.449 nM 和 419.5 μM。卡托普利的抑制效果最高，IC 值为 0.835 nM。
