Allosteric modulation of cytochrome P450 enzymes by the NADPH cytochrome P450 reductase FMN-containing domain.
NADPH-cytochrome P450 reductase
allostery
cytochrome P450
ligand binding
redox partner
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
09 2023
09 2023
Historique:
received:
23
06
2023
revised:
18
07
2023
accepted:
21
07
2023
medline:
23
10
2023
pubmed:
31
7
2023
entrez:
30
7
2023
Statut:
ppublish
Résumé
NADPH-cytochrome P450 reductase delivers electrons required by heme oxygenase, squalene monooxygenase, fatty acid desaturase, and 48 human cytochrome P450 enzymes. While conformational changes supporting reductase intramolecular electron transfer are well defined, intermolecular interactions with these targets are poorly understood, in part because of their transient association. Herein the reductase FMN domain responsible for interacting with targets was fused to the N-terminus of three drug-metabolizing and two steroidogenic cytochrome P450 enzymes to increase the probability of interaction. These artificial fusion enzymes were profiled for their ability to bind their respective substrates and inhibitors and to perform catalysis supported by cumene hydroperoxide. Comparisons with the isolated P450 enzymes revealed that even the oxidized FMN domain causes substantial and diverse effects on P450 function. The FMN domain could increase, decrease, or not affect total ligand binding and/or dissociation constants depending on both P450 enzyme and ligand. As examples, FMN domain fusion has no effect on inhibitor ketoconazole binding to CYP17A1 but substantially altered CYP21A2 binding of the same compound. FMN domain fusion to CYP21A2 resulted in differential effects dependent on whether the ligand was 17α-hydroxyprogesterone versus ketoconazole. Similar enzyme-specific effects were observed on steady-state kinetics. These observations are most consistent with FMN domain interacting with the proximal P450 surface to allosterically impact P450 ligand binding and metabolism separate from electron delivery. The variety of effects on different P450 enzymes and on the same P450 with different ligands suggests intricate and differential allosteric communication between the P450 active site and its proximal reductase-binding surface.
Identifiants
pubmed: 37517692
pii: S0021-9258(23)02140-3
doi: 10.1016/j.jbc.2023.105112
pmc: PMC10481364
pii:
doi:
Substances chimiques
CYP21A2 protein, human
EC 1.14.14.16
Flavin Mononucleotide
7N464URE7E
Ketoconazole
R9400W927I
Ligands
0
NADPH-Ferrihemoprotein Reductase
EC 1.6.2.4
Steroid 21-Hydroxylase
EC 1.14.14.16
Cytochrome P-450 Enzyme System
9035-51-2
Recombinant Fusion Proteins
0
Hydrogen Peroxide
BBX060AN9V
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
105112Subventions
Organisme : NIGMS NIH HHS
ID : R37 GM076343
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007767
Pays : United States
Informations de copyright
Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
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