Purification of glucose-6-phosphate dehydrogenase from <bold>rat</bold> (Rattus norvegicus) <bold>erythrocytes</bold> and inhibition effects of some metal ions on enzyme activity

TEMEL Y., KOÇYİĞİT Ü. M.

JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, cilt.31, sa.9, 2017 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 31 Sayı: 9
  • Basım Tarihi: 2017
  • Doi Numarası: 10.1002/jbt.21927
  • Dergi Adı: JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Anahtar Kelimeler: enzyme, erythrocyte, glucose-6-phosphate dehydrogenase (G6PD), heavy metal, purification, rat, 6-PHOSPHOGLUCONATE DEHYDROGENASE, LIVER
  • Sivas Cumhuriyet Üniversitesi Adresli: Evet

Özet

Glucose-6-phosphate dehydrogenase (G6PD) is the first enzyme on which the pentose phosphate pathway was checked. In this study, purification of a G6PD enzyme was carried out by using rat erythrocytes with a specific activity of 13.7EU/mg and a yield of 67.7 and 155.6-fold by using 2,5-ADP Sepharose-4B affinity column chromatography. For the purpose of identifying the purity of enzyme and molecular mass of the subunit, a sodium dodecyl sulfate-polyacrylamide gel electrophoresis was carried out. The molecular mass of subunit was calculated 56.5kDa approximately. Then, an investigation was carried out regarding the inhibitory effects caused by various metal ions (Fe2+, Pb2+, Cd2+, Ag+, and Zn2+) on G6PD enzyme activities, as per Beutler method at 340nm under in vitro conditions. Lineweaver-Burk diagrams were used for estimation of the IC50 and K-i values for the metals. K-i values for Pb+2, Cd+2, Ag+, and Zn+2 were 113.3, 215.2, 19.4, and 474.7M, respectively.