Electrochemical reduction of sterol-14alpha-demethylase from Mycobacterium tuberculosis (CYP51b1).
The electrochemical reduction of the heme protein sterol-14alpha-demethylase from Mycobacterium tuberculosis (CYP51b1, or further CYP51) was investigated. Direct electron transfer was demonstrated between CYP51 and graphite screen-printed electrodes modified with gold nanoparticles and with the membrane-like synthetic surfactant didodecyl dimethylammonium bromide. The formal potential of the Fe3+/Fe2+ pair, E(1/2), is equal to -273 mV (vs. Ag/AgCl). The cathodic current corresponding to the reduction of oxygen by immobilized heme protein was registered in the presence of oxygen. Addition of lanosterol, one of the substrates of the CYP51 family, to the oxygenated solution caused a concentration-dependent increase in the reduction current in voltammetric and amperometric experiments. Ketoconazole, an inhibitor of CYP51, inhibited the catalytic cathodic current in the presence of lanosterol. Electrochemical reduction of CYP51 may serve as an adequate alternative to the reconstituted system, which requires additional redox partners for the exhibition of catalytic activity of heme proteins of the cytochrome P450 superfamily.
Shumyantseva
V V
VV
Orekhovich Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow 119121, Russia. viktoria.shumyantseva@ibmc.msk.ru
Bulko
T V
TV
Kuznetsova
G P
GP
Lisitsa
A V
AV
Ponomarenko
E A
EA
Karuzina
I I
II
Archakov
A I
AI
D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000302:isolation & purification; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000302:isolation & purification; D004563:Electrochemistry; D004566:Electrodes; D006046:Gold / Q000737:chemistry; D007654:Ketoconazole / Q000494:pharmacology; D007810:Lanosterol / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D010084:Oxidation-Reduction
D001426:Bacterial Proteins; D065607:Cytochrome P-450 Enzyme Inhibitors; D007810:Lanosterol; D006046:Gold; D003577:Cytochrome P-450 Enzyme System; C501935:cytochrome P-450 CYP51, Mycobacterium tuberculosis; D007654:Ketoconazole