BACKGROUND: Interindividual variability in cytochrome P450 (CYP)-mediated xenobiotic metabolism is extensive. CYP metabolism requires two electrons, which can be donated by NADPH cytochrome P450 oxidoreductase (CYPOR) and/or cytochrome b5 (b5). Although substantial number of studies have reported on the function and effect of b5 in CYP-mediated catalysis, its mode of action is still not fully understood. OBJECTIVE: The aim of this work was to examine the effect of b5 on the activities of eight natural-occurring variants of human CYP1A2, namely, T83M, S212C, S298R, G299S, I314V, I386F, C406Y, and R456H. MATERIALS AND METHODS: An approach, as used in our former study was applied, coexpressing these polymorphic CYP1A2 variants separately with CYPOR and b5 in the bacterial cell model BTC-CYP. For each variant, 16 different activity parameters were measured, using eight different substrates. This heterogeneous data set was merged with the one of our former study (i.e. without b5) and a multivariate analysis was carried out. RESULTS: This analysis indicated that b5 seems to have the ability to affect CYP1A2 variants to behave more like the wild-type variant. This was especially the case for variant I386F, for which the presence of b5 was crucial to show activity. Variants T83M and C406Y showed considerably different activity-profiles when in the presence of b5. Furthermore, our data seem to implicate CYP1A2 residue G299 in its interaction with CYPOR and/or b5. CONCLUSION: Results indicate the ability of b5 to affect CYP1A2 variants to behave more like the wild-type variant, attenuating detrimental effects of structural mutations of these variants, seemingly through extensive allosteric effects.
BACKGROUND: Interindividual variability in cytochrome P450 (CYP)-mediated xenobiotic metabolism is extensive. CYP metabolism requires two electrons, which can be donated by NADPH cytochrome P450 oxidoreductase (CYPOR) and/or cytochrome b5 (b5). Although substantial number of studies have reported on the function and effect of b5 in CYP-mediated catalysis, its mode of action is still not fully understood. OBJECTIVE: The aim of this work was to examine the effect of b5 on the activities of eight natural-occurring variants of humanCYP1A2, namely, T83M, S212C, S298R, G299S, I314V, I386F, C406Y, and R456H. MATERIALS AND METHODS: An approach, as used in our former study was applied, coexpressing these polymorphic CYP1A2 variants separately with CYPOR and b5 in the bacterial cell model BTC-CYP. For each variant, 16 different activity parameters were measured, using eight different substrates. This heterogeneous data set was merged with the one of our former study (i.e. without b5) and a multivariate analysis was carried out. RESULTS: This analysis indicated that b5 seems to have the ability to affect CYP1A2 variants to behave more like the wild-type variant. This was especially the case for variant I386F, for which the presence of b5 was crucial to show activity. Variants T83M and C406Y showed considerably different activity-profiles when in the presence of b5. Furthermore, our data seem to implicate CYP1A2 residue G299 in its interaction with CYPOR and/or b5. CONCLUSION: Results indicate the ability of b5 to affect CYP1A2 variants to behave more like the wild-type variant, attenuating detrimental effects of structural mutations of these variants, seemingly through extensive allosteric effects.
Authors: Michael Fasullo; Julian Freedland; Nicholas St John; Cinzia Cera; Patricia Egner; Matthew Hartog; Xinxin Ding Journal: Environ Mol Mutagen Date: 2017-04-24 Impact factor: 3.216
Authors: Diana Campelo; Thomas Lautier; Philippe Urban; Francisco Esteves; Sophie Bozonnet; Gilles Truan; Michel Kranendonk Journal: Front Pharmacol Date: 2017-10-30 Impact factor: 5.810
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Authors: Barbara M A van Vugt-Lussenburg; Liliana Capinha; Jelle Reinen; Martijn Rooseboom; Michel Kranendonk; Rob C A Onderwater; Paul Jennings Journal: Chem Res Toxicol Date: 2022-06-29 Impact factor: 3.973