Modulation of direct electron transfer of cytochrome c by use of a molecularly imprinted thin film
Modulation of direct electron transfer of cytochrome c by use of a molecularly imprinted thin film
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DOI：
10.1007/s00216-013-7009-8
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发表时间：
2013-08-01
影响因子：
4.3
通讯作者：
Scheller, Frieder W.
中科院分区：
文献类型：
--
作者：
Bosserdt, Maria;Gajovic-Eichelman, Nenad;Scheller, Frieder W.
我们描述了一种分子印迹聚合物膜（MIP）的自组装单层（SAM）的巯基十一烷酸（MUA）上的黄金，其中模板细胞色素c（细胞色素c）参与直接电子转移（DET）与底层电极的顶部上的制备。为了实现DET，将不导电的聚合物膜从东莨菪亭和细胞色素c的水溶液电沉积到预先用MUA修饰的金电极的表面上。细胞色素c的电活性表面浓度为0.5pmolcm ~（-2）。在不存在MUA层的情况下，未观察到cyt c DET，并且通过在过氧化氢存在下氧化Ampliflu red评估的东莨菪碱包封的蛋白质的假过氧化物活性仅为MUA上MIP的30%。该结果表明，通过MUA-SAM对cyt c的静电吸附显著增加了电沉积步骤期间cyt c的表面浓度，并且是DET所需的生产性取向的先决条件。在用硫酸处理去除模板后，Cytc与MUA-MIP修饰电极发生重新结合，亲和常数为100，000 mol（-1）L，该值比使用荧光滴定法测定的溶液中东莨菪内酯与Cytc之间的相互作用高3倍。在肌红蛋白，溶菌酶，和牛血清白蛋白（BSA）的存在下，细胞色素C的DET揭示了MIP层抑制竞争蛋白质的作用。
We describe the preparation of a molecularly imprinted polymer film (MIP) on top of a self-assembled monolayer (SAM) of mercaptoundecanoic acid (MUA) on gold, where the template cytochrome c (cyt c) participates in direct electron transfer (DET) with the underlying electrode. To enable DET, a non-conductive polymer film is electrodeposited from an aqueous solution of scopoletin and cyt c on to the surface of a gold electrode previously modified with MUA. The electroactive surface concentration of cyt c was 0.5 pmol cm(-2). In the absence of the MUA layer, no cyt c DET was observed and the pseudo-peroxidatic activity of the scopoletin-entrapped protein, assessed via oxidation of Ampliflu red in the presence of hydrogen peroxide, was only 30 % of that for the MIP on MUA. This result indicates that electrostatic adsorption of cyt c by the MUA-SAM substantially increases the surface concentration of cyt c during the electrodeposition step, and is a prerequisite for the productive orientation required for DET. After template removal by treatment with sulfuric acid, rebinding of cyt c to the MUA-MIP-modified electrode occurred with an affinity constant of 100,000 mol(-1) L, a value three times higher than that determined by use of fluorescence titration for the interaction between scopoletin and cyt c in solution. The DET of cyt c in the presence of myoglobin, lysozyme, and bovine serum albumin (BSA) reveals that the MIP layer suppresses the effect of competing proteins.