| Autor: |
Marth G; School of Chemistry and Institute for Life Sciences, University of Southampton , Highfield, Southampton SO17 1BJ, United Kingdom., Hartley AM; School of Biosciences, Cardiff University , Cardiff CF10 3AT, United Kingdom., Reddington SC; School of Biosciences, Cardiff University , Cardiff CF10 3AT, United Kingdom., Sargisson LL; School of Chemistry and Institute for Life Sciences, University of Southampton , Highfield, Southampton SO17 1BJ, United Kingdom., Parcollet M; School of Chemistry and Institute for Life Sciences, University of Southampton , Highfield, Southampton SO17 1BJ, United Kingdom., Dunn KE; Department of Electronics, University of York , Heslington, York YO10 5DD, United Kingdom., Jones DD; School of Biosciences, Cardiff University , Cardiff CF10 3AT, United Kingdom., Stulz E; School of Chemistry and Institute for Life Sciences, University of Southampton , Highfield, Southampton SO17 1BJ, United Kingdom. |
| Abstrakt: |
We demonstrate an approach that allows attachment of single-stranded DNA (ssDNA) to a defined residue in a protein of interest (POI) so as to provide optimal and well-defined multicomponent assemblies. Using an expanded genetic code system, azido-phenylalanine (azF) was incorporated at defined residue positions in each POI; copper-free click chemistry was used to attach exactly one ssDNA at precisely defined residues. By choosing an appropriate residue, ssDNA conjugation had minimal impact on protein function, even when attached close to active sites. The protein-ssDNA conjugates were used to (i) assemble double-stranded DNA systems with optimal communication (energy transfer) between normally separate groups and (ii) generate multicomponent systems on DNA origami tiles, including those with enhanced enzyme activity when bound to the tile. Our approach allows any potential protein to be simply engineered to attach ssDNA or related biomolecules, creating conjugates for designed and highly precise multiprotein nanoscale assembly with tailored functionality. |
