Purification of Glutathione Reductase from Human Erythrocytes: Inhibition Profile of Some Anti-Epileptic Drugs

Öz

Glutathione reductase (GR) is found in the NADPH-dependent oxidoreductase family. GR has various important functions in the cell, such as protein and DNA biosynthesis, the detoxification of reactive oxygen species and free radicals. The purpose of this research was to perform the in vitro inhibition effects of anti-epileptic drugs (phenytoin, gabapentin, and primidone) on GR enzyme. In the current study, the GR enzyme was purified from human erythrocytes with a specific activity of 20.08 EU/mg protein and 2135.97-purification fold. To determine the inhibition effects of anti-epileptic drugs on GR enzyme, Lineweaver-Burk graphs were drawn for each inhibitor. Ki values and inhibition types were determined from these plotted graphs. The Ki values of drugs were found in ranging from 0.15± 0.03-5.74±1.14 mM. Phenytoin was shown the most effective inhibitor feature with a competitive inhibition type.

Anahtar Kelimeler

• anti-epileptic drugs,enzyme inhibition,enzyme purification,glutathione reductase

Glutatyon Redüktaz Enziminin İnsan Eritrositlerinden Saflaştırılması: Bazı Anti-epileptik ilaçların İnhibisyon Profili

Öz

Glutatyon redüktaz (GR), NADPH'ye bağlı oksidoredüktaz ailesinde bulunur. GR hücrede protein ve DNA biyosentezi, reaktif oksijen türlerinin ve serbest radikallerin detoksifikasyonu gibi çeşitli önemli fonksiyonlara sahiptir. Bu çalışmanın amacı, GR enzimi üzerine antiepileptik ilaçların (fenintoin, gabapentin ve pirimidon) in vitro etkilerini belirlemektir. Bu çalışmada, GR enzimi insan eritrositlerinden 20.08 EU/mg protein spesifik aktivite ve 2135.97 kat saflaştırıldı. Anti-epileptik ilaçların GR enzimi üzerindeki inhibisyon etkisini belirlemek için Lineweaver-Burk grafikleri çizildi; Ki sabiti ve inhibisyon tipleri bu çizilen grafiklerden hesaplandı. Ki değerleri 0.15± 0.03-5.74±1.14 mM aralığında bulundu. Fenintoin en etkili inhibitör özelliğini yarışmalı inhibisyon tipi ile göstermiştir.

Anahtar Kelimeler

• Anti-epileptik ilaçlar,enzim inhibisyonu,enzim saflaştırılması,glutatyon redüktaz

Kaynakça

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