Methoxy-resorufin ether as an electrochemically active biological probe for cytochrome P450 O-demethylation.
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).
Jenkins
A Toby A
AT
Department of Chemistry, University of Bath, Bath, BA2 7AY, United Kingdom. a.t.a.jenkins@bath.ac.uk
Dash
Hayley-Ann
HA
Boundy
Sam
S
Halliwell
Catherine M
CM
ffrench-Constant
Richard H
RH
D000818:Animals; D002248:Carbon Monoxide / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000145:classification / Q000166:cytology / Q000201:enzymology; D004563:Electrochemistry; D008745:Methylation; D008861:Microsomes / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009249:NADP / Q000378:metabolism; D010078:Oxazines / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D013997:Time Factors
D010078:Oxazines; D009249:NADP; C076580:7-methoxyresorufin; D002248:Carbon Monoxide; D003577:Cytochrome P-450 Enzyme System