Patole Samson, Vasilev Cvetelin, El-Zubir Osama, Wang Lin, Johnson Matthew P, Cadby Ashley J, Leggett Graham J, Hunter C Neil
Department of Chemistry , University of Sheffield , Brook Hill, Sheffield S3 7HF , UK ; Department of Molecular Biology and Biotechnology , University of Sheffield , Western Bank, Sheffield S10 2TN , UK.
Department of Molecular Biology and Biotechnology , University of Sheffield , Western Bank, Sheffield S10 2TN , UK.
Interface Focus. 2015 Aug 6;5(4):20150005. doi: 10.1098/rsfs.2015.0005.
We describe a facile approach for nanopatterning of photosynthetic light-harvesting complexes over macroscopic areas, and use optical spectroscopy to demonstrate retention of native properties by both site-specifically and non-specifically attached photosynthetic membrane proteins. A Lloyd's mirror dual-beam interferometer was used to expose self-assembled monolayers of amine-terminated alkylthiolates on gold to laser irradiation. Following exposure, photo-oxidized adsorbates were replaced by oligo(ethylene glycol) terminated thiols, and the remaining intact amine-functionalized regions were used for attachment of the major light-harvesting chlorophyll-protein complex from plants, LHCII. These amine patterns could be derivatized with nitrilotriacetic acid (NTA), so that polyhistidine-tagged bacteriochlorophyll-protein complexes from phototrophic bacteria could be attached with a defined surface orientation. By varying parameters such as the angle between the interfering beams and the laser irradiation dose, it was possible to vary the period and widths of NTA and amine-functionalized lines on the surfaces; periods varied from 1200 to 240 nm and linewidths as small as 60 nm (λ/4) were achieved. This level of control over the surface chemistry was reflected in the surface topology of the protein nanostructures imaged by atomic force microscopy; fluorescence imaging and spectral measurements demonstrated that the surface-attached proteins had retained their native functionality.
我们描述了一种在宏观区域对光合光捕获复合物进行纳米图案化的简便方法,并使用光谱学来证明通过位点特异性和非特异性附着的光合膜蛋白保留了天然特性。使用劳埃德镜双光束干涉仪将金上胺基封端的烷基硫醇盐自组装单分子层暴露于激光照射下。照射后,光氧化的吸附物被聚乙二醇封端的硫醇取代,剩余完整的胺功能化区域用于附着来自植物的主要光捕获叶绿素-蛋白质复合物LHCII。这些胺图案可以用次氮基三乙酸(NTA)衍生化,从而使来自光合细菌的多组氨酸标记的细菌叶绿素-蛋白质复合物能够以确定的表面取向附着。通过改变诸如干涉光束之间的角度和激光照射剂量等参数,可以改变表面上NTA和胺功能化线条的周期和宽度;周期从1200到240nm不等,实现了小至60nm(λ/4)的线宽。对表面化学的这种控制水平反映在通过原子力显微镜成像的蛋白质纳米结构的表面拓扑结构中;荧光成像和光谱测量表明,表面附着的蛋白质保留了其天然功能。