Mehmet ÇİFTCİ, Ümit Muhammet KOÇYİĞİT, Yusuf TEMEL, Sarkat Salim Mohammed TAHER, Mohammad Ahmad HAMZA, İbrahim Hamdi SHAFEEQ

Bazı Metal İyonlarının Japon Bıldırcın (Coturnix coturnix japonica) Karaciğer Glutatyon Redüktaz Enzimi Aktivitesi Üzerine İnhibisyon Etkilerinin Araştırılması

Bu çalışmada, bazı ağır metal iyonlarının metabolizmada önemli fonksiyonlara sahip olan glutatyon reduktaz (GR) enziminin aktivitesi üzerine inhibisyon etkileri araştırılmıştır. Bu amaçla, GR enzimi bıldırcın karaciğerinden amonyum sülfat çöktürmesi ve 2ʹ, 5ʹ-ADP Sepharose 4B afinite kromatografisi kullanılarak 19.33 EU/mg spesifik aktivite ile % 14.06 verimle 128.8 kat saflaştırılmıştır. GR enziminin saflığını kontrolü için, sodyum dodesil sülfat poliakrilamid jel elektroforezi (SDS-PAGE) yapıldı ve jelde tek bant gözlendi. GR enziminin aktivitesi, Carlberg ve Mannervik yöntemine göre 340 nm'de spektrofotometrik olarak ölçüldü. Daha sonra, GR enziminin aktivitesi üzerine farklı ağır metal iyonlarının (Co 2+, Zn 2+, Pb 2+, Fe 2+, Cd 2+, Al 3+ ve Fe 3+) inhibisyon etkileri in vitro koşullarda araştırıldı. Bıldırcın karaciğer dokusundan saflaştırılan GR enzim aktivitesinin % 50'sinin in vitro koşullarda inhibe edildiği ağır metal konsantrasyonları (IC50) değerleri belirlendi. Son olarak, bu metal iyonları için Ki değerleri Lineweaver-Burk grafiğinden hesaplandı.

Investigation of the Inhibition Effects of Some Metal Ions on Glutathione Reductase Enzyme From Japanese Quail (Coturnix coturnix japonica) Liver

In the present study, the inhibition effect of some heavy metal ions on glutathione reductase (GR) enzyme which has important functions in metabolism, was investigated. For this purpose, GR enzyme was purified with 19.33 EU/mg specific activity, a yield of 14.06% and 128.8-fold from quail liver by using ammonium sulphate precipitation and 2ʹ, 5ʹ-ADP Sepharose 4B affinity chromatography. To check the purity of GR enzyme, sodium dodecyl sulfate polyacrilamide gel electrophoresis (SDS-PAGE) was performed and single band was observed. The activity of the GR enzyme was measured spectrophotometrically at 340 nm according to the method of Carlberg and Mannervik. Then, the inhibitory effects of different heavy metal ions (Co 2+, Zn 2+, Pb 2+, Fe 2+, Cd 2+, Al 3+ and Fe 3+) on the activity of GR enzyme were examined under in vitro conditions. For the GR enzyme from quail liver tissue, heavy metal concentrations (IC50) were obtained in which 50% of GR enzyme activity was inhibited in vitro conditions. Finally, Ki values for these metal ions were calculated from the Lineweaver-Burk plots.

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