Role of Arginine 117 in Substrate Recognition by Human Cytochrome P450 2J2.

Pierre Lafite,Patrick Dansette,François André,Daniel Mansuy,Darryl Zeldin,Joan Graves

doi:10.3390/ijms19072066

Abstract

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.

Highlights

Cytochromes P450 (CYPs) form a huge superfamily of hemoproteins that play key roles in the metabolism of a large variety of xenobiotics and endogenous compounds [1]
In order to further analyze the role of arginine 117 in the binding of terfenadone derivatives, we studied the influence of residue at position 117 (R117) mutations on the inhibitory effects of a series of terfenadone derivatives, whose structures are shown in Table 2, on ebastine hydroxylation
The aforementioned results on three site-directed mutants of CYP2J2 allowed us to elucidate the role of the R117 residue on the recognition of a series of terfenadone derivatives

Summary

Introduction

Cytochromes P450 (CYPs) form a huge superfamily of hemoproteins that play key roles in the metabolism of a large variety of xenobiotics and endogenous compounds [1]. 57 genes coding for CYPs have been identified [2]. CYP2J2 is the only member of the CYP2J subfamily in humans and is the only P450 that is mainly expressed in the cardiovascular system [3]. It is assumed to be the main arachidonic acid (AA) epoxygenase in the heart as the regio- and stereoselectivities of cis-epoxyeicosatrienoic acid (EET) formation by CYP2J2 match those in the human heart [3]. Several lines of evidence suggest that CYP2J2 promotes the neoplastic phenotype of carcinoma cells and stimulates metastasis [25,26], and may represent a potential target for therapy of some human cancers [27]

Methods

Results

Conclusion