Pivotal role of P450-P450 interactions in CYP3A4 allostery

The case of α-naphthoflavone

Dmitri R. Davydov, Nadezhda Y. Davydova, Elena V. Sineva, Irina Kufareva, James R. Halpert

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

We investigated the relationship between oligomerization of CYP3A4 (cytochrome P450 3A4) and its response to ANF (α-naphthoflavone), a prototypical heterotropic activator. The addition of ANF resulted in over a 2-fold increase in the rate of CYP3A4-dependent debenzylation of 7-BFC [7-benzyloxy-4-(trifluoromethyl)coumarin] in HLM (human liver microsomes), but failed to produce activation in BD Supersomes™ or Baculosomes® containing recombinant CYP3A4 and NADPH-CPR (cytochrome P450 reductase). However, incorporation of purified CYP3A4 into Supersomes™ containing only recombinant CPR reproduced the behaviour observed with HLM. The activation in this systemwas dependent on the surface density of the enzyme. Although no activation was detectable at an L/P (lipid/P450) ratio ≥750, it reached 225% at an L/P ratio of 140. To explore the relationship between this effect and CYP3A4 oligomerization, we probed P450-P450 interactions with a new technique that employs LRET (luminescence resonance energy transfer). The amplitude of LRET in mixed oligomers of the haem protein labelled with donor and acceptor fluorophores exhibited a sigmoidal dependence on the surface density of CYP3A4 in Supersomes™. The addition of ANF eliminated this sigmoidal character and increased the degree of oligomerization at low enzyme concentrations. Therefore the mechanisms of CYP3A4 allostery with ANF involve effector-dependent modulation of P450-P450 interactions.

Original languageEnglish (US)
Pages (from-to)219-230
Number of pages12
JournalBiochemical Journal
Volume453
Issue number2
DOIs
StatePublished - Jul 15 2013

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Cytochrome P-450 CYP3A
Atrial Natriuretic Factor
Oligomerization
Chemical activation
Energy Transfer
Cardiopulmonary Resuscitation
Liver Microsomes
Luminescence
Liver
Energy transfer
Lipids
NADPH-Ferrihemoprotein Reductase
Fluorophores
Enzymes
Heme
NADP
Oligomers
Modulation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Davydov, D. R., Davydova, N. Y., Sineva, E. V., Kufareva, I., & Halpert, J. R. (2013). Pivotal role of P450-P450 interactions in CYP3A4 allostery: The case of α-naphthoflavone. Biochemical Journal, 453(2), 219-230. https://doi.org/10.1042/BJ20130398
Davydov, Dmitri R. ; Davydova, Nadezhda Y. ; Sineva, Elena V. ; Kufareva, Irina ; Halpert, James R. / Pivotal role of P450-P450 interactions in CYP3A4 allostery : The case of α-naphthoflavone. In: Biochemical Journal. 2013 ; Vol. 453, No. 2. pp. 219-230.
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Davydov, DR, Davydova, NY, Sineva, EV, Kufareva, I & Halpert, JR 2013, 'Pivotal role of P450-P450 interactions in CYP3A4 allostery: The case of α-naphthoflavone', Biochemical Journal, vol. 453, no. 2, pp. 219-230. https://doi.org/10.1042/BJ20130398

Pivotal role of P450-P450 interactions in CYP3A4 allostery : The case of α-naphthoflavone. / Davydov, Dmitri R.; Davydova, Nadezhda Y.; Sineva, Elena V.; Kufareva, Irina; Halpert, James R.

In: Biochemical Journal, Vol. 453, No. 2, 15.07.2013, p. 219-230.

Research output: Contribution to journalArticle

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