The Effect of Safranin on Glutathione Reductase and Glucose 6-Phosphate Dehydrogenase Enzymes

Abstract

The aim of this study was to assess inhibitory effect of safranin on human erythrocyte glutathione reductase GR and glucose 6-phosphate dehydrogenase G6PD . For this purpose, human erythrocyte glutathione reductase was purified 2175-fold 31.35% yield using 2’,5’-ADP Sepharose 4B affinity gel and Sephadex G-200 gel filtration chromatography and erythrocyte glucose 6-phosphate dehydrogenase was purified 8681.3-fold 24.77% yield using 2’,5’-ADP Sepharose 4B affinity gel. Safranin exhibited inhibitory effects on GR and G6PD enzymes in vitro. The IC50 values of safranin were 1.15 mM for GR and 3.22 mM for G6PD, respectively, and the Ki constants were 2.63 ± 0.13 and 4.53 ± 0.17 mM, respectively. Safranin displayed noncompetitive inhibition for GR and G6PD.

Keywords

• Safranin,glutathione reductase,glucose 6-phosphate dehydrogenase

Glutatyon Redüktaz ve Glukoz 6-Fosfat Dehidrogenaz Enzimleri Üzerine Safraninin Etkisi

Abstract

Bu çalışmanın amacı safraninin insan eritrosit glutayon redüktaz GR ve glukoz 6-fosfat dehidrogenaz G6PD üzerindeki inhibitör etkilerini değerlendirmektir. Bu amaçla, 2’,5’-ADP Sepharose 4B afinite jeli ve Sephadex G-200 jel filtrasyon kromatografisi kullanılarak insan eritrosit glutayon redüktaz 2175-kat %31.35 verim ve 2’,5’-ADP Sepharose 4B afinite jeli kullanılarak eritrosit glukoz 6-fosfat dehidrogenaz 8681.3-kat %24.77 verim ile saflaştırıldı Safranin in vitro GR ve G6PD inhibitör etkisi gösterdi. Safraninde IC50 değerleri, sırası ile GR için 1.15 mM ve G6PD için 3.22 mM, ve Ki sabitleri sırası ile 2.63 ± 0.13 ve 4.53 ± 0.17 mM oldu. Safranin GR ve G6PD için yarışmasız inhibisyon gösterdi.

Keywords

• Safranin,glutatyon redüktaz,glukoz 6-fosfat dehidrogenaz

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