Enhancing ferryl accumulation in H2O2-dependent cytochrome P450s

Jose A Amaya; Olivia M Manley; Julia C Bian; Cooper D Rutland; Nicholas Leschinsky; Steven C Ratigan; Thomas M Makris

doi:10.1016/j.jinorgbio.2023.112458

Enhancing ferryl accumulation in H₂O₂-dependent cytochrome P450s

J Inorg Biochem. 2024 Mar:252:112458. doi: 10.1016/j.jinorgbio.2023.112458. Epub 2023 Dec 20.

Authors

Jose A Amaya¹, Olivia M Manley², Julia C Bian¹, Cooper D Rutland¹, Nicholas Leschinsky¹, Steven C Ratigan¹, Thomas M Makris³

Affiliations

¹ Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208, United States of America.
² Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208, United States of America; Department of Structural and Molecular Biochemistry, North Carolina State University, Raleigh, NC 27695, United States of America.
³ Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208, United States of America; Department of Structural and Molecular Biochemistry, North Carolina State University, Raleigh, NC 27695, United States of America; Department of Chemistry, North Carolina State University, Raleigh, NC 27695, United States of America. Electronic address: tmmakris@ncsu.edu.

PMID: 38141432
DOI: 10.1016/j.jinorgbio.2023.112458

Abstract

A facile strategy is presented to enhance the accumulation of ferryl (iron(IV)-oxo) species in H₂O₂ dependent cytochrome P450s (CYPs) of the CYP152 family. We report the characterization of a highly chemoselective CYP decarboxylase from Staphylococcus aureus (OleT_SA) that is soluble at high concentrations. Examination of OleT_SA Compound I (CpdI) accumulation with a variety of fatty acid substrates reveals a dependence on resting spin-state equilibrium. Alteration of this equilibrium through targeted mutagenesis of the proximal pocket favors the high-spin form, and as a result, enhances Cpd-I accumulation to nearly stoichiometric yields.

Keywords: CYP152; Compound I; Cytochrome P450; Ferryl; Peroxygenase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cytochrome P-450 Enzyme System* / chemistry
Fatty Acids / chemistry
Hydrogen Peroxide*

Substances

Cytochrome P-450 Enzyme System
Hydrogen Peroxide
Fatty Acids