Shumyantseva Victoria V, Bulko Tatiana V, Samenkova Natalia F, Kuznetsova Galina P, Usanov Sergei A, Schulze Holger, Bachmann Till T, Schmid Rolf D, Archakov Alexander I
Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Pogodinskaya Street, 10, Moscow 119121, Russia.
J Inorg Biochem. 2006 Aug;100(8):1353-7. doi: 10.1016/j.jinorgbio.2006.03.007. Epub 2006 Apr 25.
New approach to the electrochemical reduction of cytochromes P450 (P450s, CYPs) at electrodes chemically modified with appropriate substrates for P450s ("reverse" electrodes) was proposed. The method is based on the analysis of cyclic voltammograms, square-wave voltammograms and amperograms with subsequent determination of electrochemical characteristics such as catalytic current and redox potential. The sensitivity of proposed method is 0.2-1 nmol P450/electrode. The changes of maximal current and of redox potentials in square-wave voltammograms as well as the changes of catalytic current in amperometric experiments proved to be informative and reliable. Planar regime of screen-printed electrodes (strip-type sensors) enabled to utilise 20-60 microl of electrolyte volume. The enzyme-substrate pairs P450 2B4/benzphetamine and P450scc/cholesterol were investigated. Electrochemical parameters of electrodes with unspecific P450 substrates differed considerably from electrodes with appropriate substrates.
提出了一种在经细胞色素P450（P450s，CYPs）合适底物化学修饰的电极（“反向”电极）上对细胞色素P450进行电化学还原的新方法。该方法基于对循环伏安图、方波伏安图和电流图的分析，随后测定诸如催化电流和氧化还原电位等电化学特性。所提出方法的灵敏度为0.2 - 1 nmol P450/电极。方波伏安图中最大电流和氧化还原电位的变化以及安培实验中催化电流的变化被证明是信息丰富且可靠的。丝网印刷电极（条型传感器）的平面模式能够使用20 - 60微升的电解液体积。研究了酶 - 底物对P450 2B4/苄非他明和P450scc/胆固醇。具有非特异性P450底物的电极的电化学参数与具有合适底物的电极有很大差异。
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