Stoichiometry of 7‐ethoxycoumarin metabolism by cytochrome P450 2B1 wild‐type and five active‐site mutants

FEBS Letters

Fang, Xiaojun¹  Halpert, James R¹  Kobayashi, Yasuna¹ 

[1] Department of Pharmacology and Toxicology, College of Pharmacy, The University of Arizona, Tucson, AZ 85721, USA

关键词: Stoichiometry;    Cytochrome P450 2B1;    Active-site mutant;    Uncoupling;    7-Ethoxycoumarin;    Hydrogen peroxide;    P450;    cytochrome P450;    CHAPS;    (3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate;    EDTA;    ethylenediaminetetraacetic acid;    HEPES;    N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    DLPC;    dilauroyl-l-3-phosphatidylcholine;    reductase;    NADPH-cytochrome P450 reductase;   

DOI  :  10.1016/S0014-5793(97)01173-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Recombinant P450 2B1 wild-type and the active-site mutants I114V, F206L, V363A, V363L, and G478S were purified and studied. The efficiency of coupling of reducing equivalents to 7-hydroxycoumarin formation was decreased for all the mutants except I114V. Uncoupling to H₂O was increased for F206L, V363A, and G478S, decreased for V363L, and unchanged for I114V. Uncoupling to H₂O₂ was increased for V363L and decreased for I114V, F206L, and V363A. The findings from this study provide firm biochemical evidence that residues 206, 363, and 478 comprise part of the substrate binding site of P450 2B1.

【 授权许可】

Unknown