Site-specific Protein Photochemical Covalent Attachment to Carbon Nanotube Side Walls and its Electronic Impact on Single Molecule Function.
Site-specific Protein Photochemical Covalent Attachment to Carbon Nanotube Side Walls and its Electronic Impact on Single Molecule Function.
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DOI：
10.1021/acs.bioconjchem.9b00719
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发表时间：
2019-11
影响因子：
4.7
通讯作者：
Suzanne Thomas;David Jamieson;R. Gwyther;B. Bowen;A. Beachey;H. Worthy;J. Macdonald;M. Elliott;Oliver Kieran Castell;Darran Dafydd Jones
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作者：
Suzanne Thomas;David Jamieson;R. Gwyther;B. Bowen;A. Beachey;H. Worthy;J. Macdonald;M. Elliott;Oliver Kieran Castell;Darran Dafydd Jones
蛋白质与碳基纳米材料（如纳米管）的功能集成在新兴电子和光电子应用中具有很大的前景。对蛋白质附着的控制是一致和有用的设备制造的主要挑战，特别是当利用单/少数分子特性时。在这里，我们利用遗传编码的苯叠氮化物光化学来定义四种不同蛋白质（包括荧光蛋白GFP和-内酰胺酶结合蛋白（BBP））与碳纳米管侧壁的直接共价附着。AFM结果表明，附着体上BBP仍能识别并结合其他蛋白组分。单分子荧光显示，附着在SWCNTs上的功能得以保留，并且在荧光强度和抗光漂白能力方面对GFP有反馈；绿色荧光蛋白在附着上的荧光持续时间更长。在对GFP功能的电子影响方面，附着位点被证明是重要的，离发色团最远的附着位点对荧光的影响更大。我们的方法为生成蛋白质-碳纳米管杂交生物偶联物提供了一种通用的方法。它可以很容易地应用于任何选择的蛋白质；通过基因诱变，简单地将叠氮化苯放置在不同的残基上，就可以很容易地改变与碳纳米管的附着位置和界面特征。因此，我们的方法将通过改变蛋白质附着位置来实现一致的构建和调节功能偶联。
Functional integration of proteins with carbon-based nanomaterials such as nanotubes holds great promise in emerging electronic and optoelectronic applications. Control over protein attachment poses a major challenge for consistent and useful device fabrication, especially when utilizing single/few molecule properties. Here, we exploit genetically encoded phenyl azide photochemistry to define the direct covalent attachment of four different proteins, including the fluorescent protein GFP and a -lactamase binding protein (BBP), to carbon nanotube side walls. AFM showed that on attachment BBP could still recognize and bind additional protein components. Single molecule fluorescence revealed that on attachment to SWCNTs function was retained and there was feedback to GFP in terms of fluorescence intensity and improved resistance to photobleaching; GFP is fluorescent for much longer on attachment. The site of attachment proved important in terms of electronic impact on GFP function, with the attachment site furthest from the chromophore having the larger effect on fluorescence. Our approach provides a versatile and general method for generating intimate protein-CNT hybrid bioconjugates. It can be potentially applied easily to any protein of choice; attachment position and thus interface characteristics with the CNT can easily be changed by simply placing the phenyl azide chemistry at different residues by gene mutagenesis. Thus, our approach will allow consistent construction and modulate functional coupling through changing the protein attachment position.