Functional characterization of eight human CYP1A2 variants: The role of cytochrome b5

Bernardo B. Palma, Marta Silva E Sousa, Phillipe Urban, José Rueff, Michel Kranendonk

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)41-52
Number of pages12
JournalPharmacogenetics And Genomics
Volume23
Issue number2
DOIs
Publication statusPublished - 1 Feb 2013

Fingerprint

Cytochromes b5
Cytochrome P-450 CYP1A2
Cytochrome P-450 Enzyme System
Oxidoreductases
NADPH-Ferrihemoprotein Reductase
Xenobiotics
Catalysis
Multivariate Analysis
Electrons
Mutation

Keywords

  • CYP1A2
  • cytochrome b5
  • cytochrome P450
  • pharmacogenomics
  • pharmacokinetics
  • polymorphisms

Cite this

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title = "Functional characterization of eight human CYP1A2 variants: The role of cytochrome b5",
abstract = "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.",
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Functional characterization of eight human CYP1A2 variants : The role of cytochrome b5. / Palma, Bernardo B.; Silva E Sousa, Marta; Urban, Phillipe; Rueff, José; Kranendonk, Michel.

In: Pharmacogenetics And Genomics, Vol. 23, No. 2, 01.02.2013, p. 41-52.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Functional characterization of eight human CYP1A2 variants

T2 - The role of cytochrome b5

AU - Palma, Bernardo B.

AU - Silva E Sousa, Marta

AU - Urban, Phillipe

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AU - Kranendonk, Michel

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N2 - 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.

AB - 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.

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