Enhanced purification protocol for the angiotensin-converting enzyme from bovine systems and investigation of the in vitro effect of some active substances

Karahan F., Türkoğlu V.

CHEMICO-BIOLOGICAL INTERACTIONS, cilt.347, 2021 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 347
  • Basım Tarihi: 2021
  • Doi Numarası: 10.1016/j.cbi.2021.109604
  • Dergi Adı: CHEMICO-BIOLOGICAL INTERACTIONS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, BIOSIS, CAB Abstracts, Chemical Abstracts Core, EMBASE, MEDLINE, Veterinary Science Database
  • Anahtar Kelimeler: Angiotensin-converting enzyme, Beta-blockers, Diuretic, Inhibition, Purification, BETA-BLOCKERS, BLOOD-PRESSURE, LUNG, HYPERTENSION, DIURETICS, ATENOLOL, ASSAY, ACE, LISINOPRIL, PEPTIDES
  • Van Yüzüncü Yıl Üniversitesi Adresli: Evet

Özet

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 degrees 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. IC50 values of captopril, fosinopril, lisinopril, and triamterene from the graphical equation were calculated as 0.835 nM, 1.159 IrM, 4.085 nM, and 227 IrM, respectively. The inhibition type and Ki values of these compounds were determined from Lineweaver-Burk plots. Captopril, fosinopril, lisinopril, and triamterene demonstrated a non-competitive inhibition effect on ACE activity. Ki constants were found as 1.057 nM, 1.675 IrM, 6.449 nM, and 419.5 IrM, respectively. Captopril indicated the highest inhibitor effect with an IC50 value of 0.835 nM.