Methoxy-resorufin ether as an electrochemically active biological probe for cytochrome P450 O-demethylation

• Published in: Bioelectrochemistry (Amsterdam, Netherlands) Vol. 68; no. 1; pp. 67 - 71
• Main Authors: Jenkins, A. Toby A., Dash, Hayley-Ann, Boundy, Sam, Halliwell, Catherine M., ffrench-Constant, Richard H.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 2006
• Subjects: Animals; Carbon Monoxide - pharmacology; CYP6G1; Cytochrome P-450 Enzyme System - analysis; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - metabolism; Cytochrome P450; Drosophila melanogaster; Drosophila melanogaster - classification; Drosophila melanogaster - cytology; Drosophila melanogaster - enzymology; Electrochemistry; Methylation; Microsomes - chemistry; Microsomes - metabolism; Molecular Structure; MROD; NADP - metabolism; Oxazines - chemistry; Resorufin; Spectrometry, Fluorescence; Substrate Specificity; Time Factors; Electrochemistry; Cytochrome P450; CYP6G1; MROD; Resorufin
• ISSN: 1567-5394
• DOI: 10.1016/j.bioelechem.2005.04.002

This paper describes the utilisation of methoxy-resorufin ether as an electrochemical probe for studying cytochrome
P450 CYP6G1. Methoxy-resorufin ether is well established as a versatile substrate for cytochrome
P450, as its demethylated product, resorufin, is a fluorophore. We show that in addition to these established properties, methoxy-resorufin ether also exhibits reversible two electron transfer on glassy carbon and edge plane graphite electrodes. Cyclic voltammetry measurements and differential pulse voltammetry measurements show that methoxy-resorufin ether can be easily detected at low concentrations (down to 200 nM) in a conventional three electrode electrochemical cell. These properties of methoxy-resorufin ether mean that it could be used as an electrochemical probe, to follow the rate of its demethylation by CYP6G1. We show that electrochemical measurements could discriminate between the enzyme activity of protein microsomes taken from two strains of
Drosophila melanogaster (fruit fly).