Purification of cytochrome P450 2C9.1, 2C9.3 and 2C9.30 expressed in and their activities toward arachidonic acid

前川 京子*; 松澤 由美子*; 安達 基泰; 黒木 良太; 斎藤 嘉朗*

no journal, , 

Cytochrome P450 2C9 (CYP2C9) is a polymorphic enzyme responsible for the oxidative metabolism of up to 15% of the drugs that undergo phase I metabolism. We previously reported that and , which were observed at allele frequencies of about 3% and 0.2% in Japanese, respectively. This study was aimed to access their catalytic activities toward endogenous arachidonic acid . The N-terminal modified and C-terminal His-tagged wild-type CYP2C9, CYP2C93 and CYP2C930 were expressed in Rosetta2 cells. After cells were lysed, proteins were extracted from membranes using Cymal-5 and purified using Ni-affinity and CM-Sepharose ion exchange columns. Both variants exhibited significantly lower Luciferein-H hydroxylation activities than the wild-type. Arachidonic acid were metabolized into bioactive lipids such as 14,15-epoxyeicosatrienoic acid and 19-hydroxyeicosatetraenoic acid, but their production by variant enzymes was much lower than those by the wild-type.