Bazı metal iyonlarının tavuk böbreğinden saflaştırılan glutatyon redüktaz enzimi üzerine in vitro etkilerinin araştırılması

Glutayon redüktaz enzimi tavuk böbreğinden saflaştırıldı ve bazı metal iyonlarının bu enzim aktivitesi üzerine in vitro etkileri araştırıldı. Saflaştırma prosedürü üç basamaktan oluştu; homojenatın hazırlanması, amonyum sülfat çöktürmesi ve affinite kromatografisi. Bu üç basamak sonucunda 8,595 EÜ/mg protein spesifik aktivitesine sahip enzim ~ % 57 verimle 369 kat saflaştırıldı. Enzimin saflık kontrolü sodyum dodesil sülfat-poliakrilamid jel elektroforezi (SDS-PAGE) ile yapıldı ve bazı metal iyonlarının (Ni+2, Zn+2, Pb+2, Hg+2, Ag+ and Al+3) glutatyon redüktaz enzimi üzerindeki inhibisyon etkisi araştırıldı. Metal iyonları için Ki ve IC50 değerleri Lineweaver-Burk ve % vs [I] aktivite grafikleriyle belirlendi. Ni+2,  Hg+2 ve Ag+ yarışmasız inhibisyon gösterirken, Zn+2 ve Pb+2 yarı yarışmalı inhibisyon gösterdi. Diğer metal iyonlarının aksine Al+3 enzim üzerinde aktivatör etkisi gösterdi.

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