Role of Arginine 117 in Substrate Recognition by Human Cytochrome P450 2J2
Résumé
The influence of Arginine 117 of human cytochrome P450 2J2 in the recognition of ebastine and a series of terfenadone derivatives was studied by site-directed mutagenesis. R117K, R117E, and R117L mutants were produced, and the behavior of these mutants in the hydroxylation of ebastine and terfenadone derivatives was compared to that of wild-type CYP2J2. The data clearly showed the importance of the formation of a hydrogen bond between R117 and the keto group of these substrates. The data were interpreted on the basis of 3D homology models of the mutants and of dynamic docking of the substrates in their active site. These modeling studies also suggested the existence of a R117-E222 salt bridge between helices B' and F that would be important for maintaining the overall folding of CYP2J2.
Mots clés
cyp2j2
arachidonic-acid metabolism
inhibitor
docking
in-vitro
epoxyeicosatrienoic acids
epoxygenase-derived eicosanoids
homology modeling
human intestinal microsomes
human cyp2j2
human cytochrome P450
human liver-microsomes
molecular-cloning
mutants
mutation-induced dysfunction
potential functional-significance
regioselectivity