In vitro effect of oxidized and reduced glutathione peptides on angiotensin converting enzyme purified from human plasma


Basi Z., Türkoğlu V.

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, cilt.1104, ss.190-195, 2019 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 1104
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1016/j.jchromb.2018.11.023
  • Dergi Adı: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.190-195
  • Anahtar Kelimeler: Angiotensin converting enzyme (ACE), Purification, Oxidized glutathione (GSSG), Reduced glutathione (GSH), Inhibition, Activation, Lisinopril, INHIBITORY PEPTIDES, OXIDATIVE STRESS, MOLECULAR DOCKING, PURIFICATION, HYPERTENSION, ACE, PROTEIN, BLOOD, ASSAY, LUNG
  • Van Yüzüncü Yıl Üniversitesi Adresli: Evet

Özet

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 50 values for GSH and lisinopril were determined to be 16.2 mu M 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-i constants for GSH and lisinopril were calculated as 11.7 mu M and 0.662 nM, respectively.