Site-Specific Protein Photochemical Covalent Attachment to Carbon Nanotube Side Walls and Its Electronic Impact on Single Molecule Function
| Autor: | W. David Jamieson, Harley L. Worthy, Benjamin J. Bowen, Rebecca E. A. Gwyther, Adam Beachey, Martin Elliott, Suzanne K. Thomas, Oliver Kieran Castell, J. Emyr Macdonald, D. Dafydd Jones |
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| Rok vydání: | 2019 |
| Předmět: |
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Molecular Protein Conformation Green Fluorescent Proteins Biomedical Engineering Pharmaceutical Science Electrons Bioengineering 02 engineering and technology Carbon nanotube Photochemistry 01 natural sciences Green fluorescent protein law.invention chemistry.chemical_compound law Phenyl azide Pharmacology Binding Sites Nanotubes Carbon 010405 organic chemistry Binding protein Organic Chemistry Photochemical Processes 021001 nanoscience & nanotechnology Single-molecule experiment Fluorescence Photobleaching 0104 chemical sciences chemistry Covalent bond 0210 nano-technology Biotechnology |
| Zdroj: | Bioconjugate Chemistry. 31:584-594 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/acs.bioconjchem.9b00719 |
| Popis: | 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 β-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 to any protein of choice; the 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. |
| Databáze: | OpenAIRE |
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