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

Year 2018, Volume: 39 Issue: 3, 679 - 687, 30.09.2018

Yusuf Temel Sarkat Salim Mohammed Taher Mohammad Ahmad Hamza İbrahim Hamdi Shafeeq Ümit Muhammet Koçyiğit Mehmet Çiftçi

https://doi.org/10.17776/csj.414680

Cited By: 5

Abstract

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, K_i values for
these metal ions were calculated from the Lineweaver-Burk plots.

Keywords

Glutathion Reductase, Enzyme, Purification, Quail, Inhibition, Liver

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

Abstract

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.

Keywords

Glutatyon Redüktaz, Enzim, Saflaştırma, İnhibisyon, Bıldırcın, Karaciğer

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