Biased cytochrome P450-mediated metabolism via small-molecule ligands binding P450 oxidoreductase

Nat Commun. 2021 Apr 15;12(1):2260. doi: 10.1038/s41467-021-22562-w.

Abstract

Metabolic control is mediated by the dynamic assemblies and function of multiple redox enzymes. A key element in these assemblies, the P450 oxidoreductase (POR), donates electrons and selectively activates numerous (>50 in humans and >300 in plants) cytochromes P450 (CYPs) controlling metabolism of drugs, steroids and xenobiotics in humans and natural product biosynthesis in plants. The mechanisms underlying POR-mediated CYP metabolism remain poorly understood and to date no ligand binding has been described to regulate the specificity of POR. Here, using a combination of computational modeling and functional assays, we identify ligands that dock on POR and bias its specificity towards CYP redox partners, across mammal and plant kingdom. Single molecule FRET studies reveal ligand binding to alter POR conformational sampling, which results in biased activation of metabolic cascades in whole cell assays. We propose the model of biased metabolism, a mechanism akin to biased signaling of GPCRs, where ligand binding on POR stabilizes different conformational states that are linked to distinct metabolic outcomes. Biased metabolism may allow designing pathway-specific therapeutics or personalized food suppressing undesired, disease-related, metabolic pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aromatase / metabolism
  • Cell Line
  • Cytochrome P-450 Enzyme System / genetics
  • Cytochrome P-450 Enzyme System / isolation & purification
  • Cytochrome P-450 Enzyme System / metabolism*
  • Enzyme Assays
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Ligands*
  • Liposomes / metabolism
  • Metabolic Networks and Pathways*
  • Molecular Docking Simulation
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Single Molecule Imaging
  • Steroid 17-alpha-Hydroxylase / metabolism
  • Steroid 21-Hydroxylase / metabolism
  • Substrate Specificity

Substances

  • Ligands
  • Liposomes
  • POR protein, human
  • Recombinant Proteins
  • Cytochrome P-450 Enzyme System
  • Aromatase
  • CYP19A1 protein, human
  • CYP21A2 protein, human
  • Steroid 21-Hydroxylase
  • CYP17A1 protein, human
  • Steroid 17-alpha-Hydroxylase