Conformational change detection in nonmetal proteins by direct electrochemical oxidation using diamond electrodes

Masanobu Chiku, Jin Nakamura, Akira Fujishima, Yasuaki Einaga

研究成果: Article査読

55 被引用数 (Scopus)

抄録

In this report, we established a new electrochemical method for the detection of conformational changes in large, non-metalloproteins such as bovine serum albumin, using flow injection analysis coupled with hydrogen-terminated, boron-doped diamond electrodes. The oxidation current was used as a signal reporter in the monitoring of urea-induced BSA denaturation. In the denatured state at high urea concentrations, the electrochemical signal increased, and the amperometric responses for the oxidation potential at 1300 mV were consistent with the results of conventional methods of denaturation monitoring using fluorescence spectroscopy. The oxidation involved at least five redox-active species (cysteine, tryptophan, tyrosine, methionine, and disulfide bonds). Furthermore, the method also showed high sensitivity for quantitative analysis of protein. A linear dynamic in the concentration range 50-400 μg/mL (r 2 = 0.977) with a lower detection limit of 190 ng/mL was achieved for BSA. Direct electrochemical detection of conformation changes of proteins using BDD electrodes can be performed with advantages in terms of simplicity and sensitivity.

本文言語English
ページ(範囲)5783-5787
ページ数5
ジャーナルAnalytical Chemistry
80
15
DOI
出版ステータスPublished - 2008 8 1

ASJC Scopus subject areas

  • 分析化学

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