Purification and characterization of glutathione reductase from sheep liver

ULUSU G., ERAT M., CIFTCI M., Sakiroglu H., BAKAN E.

TURKISH JOURNAL OF VETERINARY & ANIMAL SCIENCES, cilt.29, sa.5, ss.1109-1117, 2005 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 29 Sayı: 5
  • Basım Tarihi: 2005
  • Dergi Adı: TURKISH JOURNAL OF VETERINARY & ANIMAL SCIENCES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.1109-1117
  • Anahtar Kelimeler: glutathione reductase, purification, characterization, sheep, liver, AFFINITY-CHROMATOGRAPHY, RAT-LIVER, ERYTHROCYTES, ENZYME, PROTEINS, BINDING
  • Atatürk Üniversitesi Adresli: Evet

Özet

Glutathione reductase (Glutathione: NADP+ oxidoreductase, EC 1.8.1.7; GR) was purified from sheep liver. The purification included 4 steps: preparation of homogenate, ammonium sulfate fractionation, 2', 5'-ADP Sepharose-4B affinity chromatography, and gel filtration chromatography. GR was obtained with a yield of 14.1% having a specific activity of 47.27 EU/mg proteins. Optimal pH, stable pH, and optimal temperature were 8.0, 5.5, and 60 degrees C, respectively. K-M and V-max values for NADPH and GSSG substrates were 0.0258 and 0.0239 mM, and 0.266 and 0.255 EU/ml, respectively. The overall purification was about 1477-fold for liver GR. The enzyme activity was measured spectrophotometrically at 340 nm. In addition, K-j values of 4.367 and 0.4055 mM were determined by means of Lineweaver-Burk graphs for NADP(+) and GSH, respectively.