A facilitated electron transfer of copper-zinc superoxide dismutase (SOD) based on a cysteine-bridged SOD electrode

被引:75
作者
Tian, Y
Shioda, M
Kasahara, S
Okajima, T
Mao, LQ
Hisabori, T
Ohsaka, T
机构
[1] Tokyo Inst Technol, Interdisciplinary Grad Sch Sci & Engn, Dept Elect Chem, Midori Ku, Yokohama, Kanagawa 2268502, Japan
[2] Tokyo Inst Technol, Resources Utilizat Res Lab, Midori Ku, Yokohama, Kanagawa 2268503, Japan
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 2002年 / 1569卷 / 1-3期
关键词
superoxide dismutase (SOD); self-assembled monolayer (SAM); cysteine; redox rcaction; electron transfer; copper zinc;
D O I
10.1016/S0304-4165(01)00246-X
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The direct electrochemical redox reaction of bovine erythrocyte copper-zinc superoxide dismutase (Cu2Zn2SOD) was clearly observed at a gold electrode modified with a self-assembled monolayer (SAM) of cysteine in phosphate buffer solution containing SOD, although its reaction could not be observed at the bare electrode. In this case, SOD was found to be stably confined on the SAM of cysteine and the redox response could be observed even when the cysteine-SAM electrode used in the SOD solution was transferred to the pure electrolyte solution containing no SOD, suggesting the permanent binding of SOD via the SAM of cysteine on the electrode surface. The electrode reaction of the SOD confined on the cysteine-SAM electrode was found to be quasi-reversible with the formal potential of 65+/-3 mV vs. Ag/ AgCI and its kinetic parameters were estimated: the electron transfer rate constant k(s), is 1.2+/-0.2 s(-1) and the anodic (alpha(a)) and cathodic (alpha(c)) transfer coefficients are 0.39+/-0.02 and 0.61+/-0.02, respectively. The assignment of the redox peak of SOD at the cysteinc-SAM modified electrode could be sufficiently carried out using the native SOD (Cu2Zn2SOD), its Cu- or Zn-free derivatives (E2Zn2SOD and Cu2E2SOD. E designates an empty site) and the SOD reconstituted from E2Zn2SOD and CU2+. The Cu complex moiety, the active site for the enzymatic dismutation of the superoxide ion, was characterized to be also the electroactive site of SOD. In addition, we found that the SOD confined on the electrode can be expected to possess its inherent enzymatic activity for dismutation of the superoxide ion. (C) 2002 Elsevier Science B.V. All rights reserved.
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页码:151 / 158
页数:8
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