Orientated binding of photosynthetic reaction centers on gold using Ni-NTA self-assembled monolayers
Orientated binding of photosynthetic reaction centers on gold using Ni-NTA self-assembled monolayers
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DOI：
10.1016/j.bios.2003.12.034
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发表时间：
2004-07-15
影响因子：
12.6
通讯作者：
Lebedev, N
中科院分区：
文献类型：
--
作者：
Trammell, SA;Wang, LY;Lebedev, N
光合反应中心与无机表面的偶联对于蛋白质间电子传递机制的研究以及光传感器和化学传感器的构建具有重要意义。在这里，我们表明，可以将来自球形红细菌的RC固定在金表面上，通过在M亚基的C末端使用基因工程多组氨酸标签（His（7））和Ni-NTA终止的自组装单层（SAM），RC主要供体朝向基底。在电子受体泛醌-10的存在下，该RC电极的照射产生阴极光电流。光电流的作用光谱与RC的吸收光谱一致，并且响应于除草剂阿特拉津的光电流减小，证实RC是光响应的主要来源。通过咪唑破坏Ni-NTA-RC键导致光电流降低约80%，表明大部分光活性蛋白通过接头特异性结合到电极。(C)2004 Elsevier B. V.保留所有权利。
Coupling of photosynthetic reaction centers (RCs) with inorganic surfaces is attractive for the identification of the mechanisms of interprotein electron transfer (ET) and for possible applications in construction of photo- and chemosensors. Here we show that RCs from Rhodobacter sphaeroides can be immobilized on gold surfaces with the RC primary donor looking towards the substrate by using a genetically engineered poly-histidine tag (His(7)) at the C-terminal end of the M-subunit and a Ni-NTA terminated self-assembled monolayer (SAM). In the presence of an electron acceptor, ubiquinone-10, illumination of this RC electrode generates a cathodic photocurrent. The action spectrum of the photocurrent coincides with the absorption spectrum of RC and the photocurrent decreases in response to the herbicide, atrazine, confirming that the RC is the primary source of the photoresponse. Disruption of the Ni-NTA-RC bond by imidazole leads to about 80% reduction of the photocurrent indicating that most of the photoactive protein is specifically bound to the electrode through the linker. (C) 2004 Elsevier B.V. All rights reserved.