Controlling the formation of a monolayer of cytochrome P450 reductase onto Au surfaces
Controlling the formation of a monolayer of cytochrome P450 reductase onto Au surfaces
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DOI：
10.1103/physreve.86.011903
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发表时间：
2012-07-03
影响因子：
2.4
通讯作者：
Weightman, P.
中科院分区：
文献类型：
--
作者：
Convery, J. H.;Smith, C. I.;Weightman, P.
用具有耗散、原子力显微镜和反射各向异性光谱(RAS)的石英晶体微天平测定了人细胞色素P450还原酶在Au(110)/电解液界面上形成突变形式(P499C)单层和双层的必要条件。分子通过Au-S键吸附，并在Au(110)衬底上呈有序结构，其中贡献RAS信号的偶极子的光轴大致沿Au(110)衬底的光轴对准。溶液中分子的吸收光谱和被吸附的单分子膜的RAS谱之间的差异归因于在光谱的低能区贡献的偶极子取向的表面有序性、异四氮杂环长轴表面的大致垂直取向以及芳香氨基酸中偶极子的分子结构没有任何择优取向。我们的研究为将生物大分子固定到金表面提供了重要的原理证据。这开启了RAS对生物大分子动力学的详细研究，在与蛋白质动力学耦合的生物氧化还原化学研究中具有广泛的应用。
The conditions necessary for the formation of a monolayer and a bilayer of a mutated form (P499C) of human cytochrome P450 reductase on a Au(110)/electrolyte interface have been determined using a quartz crystal microbalance with dissipation, atomic force microscopy, and reflection anisotropy spectroscopy (RAS). The molecules adsorb through a Au-S linkage and, for the monolayer, adopt an ordered structure on the Au(110) substrate in which the optical axes of the dipoles contributing to the RAS signal are aligned roughly along the optical axes of the Au(110) substrate. Differences between the absorption spectrum of the molecules in a solution and the RAS profile of the adsorbed monolayer are attributed to surface order in the orientation of dipoles that contribute in the low energy region of the spectrum, a roughly vertical orientation on the surface of the long axes of the isoalloxazine rings and the lack of any preferred orientation in the molecular structure of the dipoles in the aromatic amino acids. Our studies establish an important proof of principle for immobilizing large biological macromolecules to gold surfaces. This opens up detailed studies of the dynamics of biological macromolecules by RAS, which have general applications in studies of biological redox chemistry that are coupled to protein dynamics.