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Precision templated bottom-up multiprotein nanoassembly through defined click chemistry linkage to DNA

Marth, Gabriella, Hartley, Andrew M., Reddington, Samuel C., Sargisson, Lauren L., Parcollet, Marlène, Dunn, Katherine E., Jones, D. Dafydd ORCID: and Stulz, Eugen 2017. Precision templated bottom-up multiprotein nanoassembly through defined click chemistry linkage to DNA. ACS Nano 11 (5) , pp. 5003-5010. 10.1021/acsnano.7b01711

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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.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Subjects: Q Science > QH Natural history > QH426 Genetics
Uncontrolled Keywords: copper-free click chemistry; DNA nanotechnology; energy transfer; expanded genetic code; origami; precision assembly; protein engineering
Additional Information: This is an open access article under the terms of the CC-BY license.
Publisher: American Chemical Society
ISSN: 1936-0851
Date of First Compliant Deposit: 20 June 2017
Date of Acceptance: 17 April 2017
Last Modified: 05 May 2023 16:13

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