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Functional characterization and tissue expression of marmoset cytochrome P450 2E1
Author(s) -
Uehara Shotaro,
Uno Yasuhiro,
Tomioka Etsuko,
Inoue Takashi,
Sasaki Erika,
Yamazaki Hiroshi
Publication year - 2017
Publication title -
biopharmaceutics and drug disposition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.419
H-Index - 58
eISSN - 1099-081X
pISSN - 0142-2782
DOI - 10.1002/bdd.2080
Subject(s) - marmoset , callithrix , cytochrome p450 , biology , complementary dna , enzyme , cyp2e1 , hydroxylation , microbiology and biotechnology , biochemistry , gene , paleontology
Common marmosets ( Callithrix jacchus ) have attracted increasing attention as a useful small non‐human primate model in preclinical research. However, studies on marmoset cytochrome P450 (P450) 2E enzyme have scarcely been conducted. In this study, the full‐length cDNA encoding P450 2E1 enzyme was isolated from marmoset livers by reverse transcription (RT)‐polymerase chain reaction (PCR). Marmoset P450 2E1 amino acid sequences were highly identical (>88%) to those of cynomolgus monkey and human P450 2E1 enzymes. Phylogenetic analysis indicated a close evolutionary relationship among marmoset, cynomolgus monkey, and human P450 2E1 enzymes. The tissue expression pattern analyzed by real‐time RT‐PCR and immunoblotting demonstrated that marmoset P450 2E1 mRNA and proteins were predominantly expressed in livers. Marmoset P450 2E1 enzyme heterologously expressed in Escherichia coli catalyzed the hydroxylation of p ‐nitrophenol, chlorzoxazone, and theophylline, similar to cynomolgus monkey and human P450 2E1 enzymes. By kinetic analyses, those P450 2E1 enzymes catalyzed p ‐nitrophenol hydroxylation with similar affinities and relatively high intrinsic clearance efficiencies. These results indicated that tissue distribution and enzyme‐substrate specificity of marmoset P450 2E1 were similar to cynomolgus monkey and human P450 2E1 enzymes, suggesting that marmosets are a suitable primate model for P450 2E1‐dependent drug and xenobiotic metabolism.