Mediated spectroelectrochemical titration of proteins for redox potential measurements by a separator-less one-compartment bulk electrolysis method

被引：59

作者：

Tsujimura, S ^{[1
]}

Kuriyama, A ^{[1
]}

Fujieda, N ^{[1
]}

Kano, K ^{[1
]}

Ikeda, T ^{[1
]}

机构：

[1] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Sakyo Ku, Kyoto 6068502, Japan

来源：

ANALYTICAL BIOCHEMISTRY | 2005年 / 337卷 / 02期

关键词：

redox potential; redox protein; spectroelectrochemistry; bulk electrolysis; mediator;

D O I：

10.1016/j.ab.2004.11.017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

One-compartment bulk electrolysis and simultaneous spectroscopic measurements are realized in a conventional spectroscopic cuvette without separator by using a mesh-type working electrode with extremely large surface area and a wire-type counter electrode with very small surface area. Spectrophotometric monitoring revealed complete electrolysis in a first-order kinetics. This technique was applied to mediated titration of cytochrome c and bilirubin oxidase for determining their redox potentials. Kinetics for the solution redox reaction between protein and mediator is described. The subtraction of spectral background due to mediator adsorption is very easy because of high reproducibility. The experiments can be done under completely anaerobic conditions. Low-absorbance protein samples (of low concentrations or small absorption coefficients) and hydrophobic proteins (such as membrane-bound proteins) are acceptable for measurements. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：325 / 331

页数：7

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