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

Precision templated bottom-up multiprotein nanoassembly through defined click chemistry linkage to DNA
Precision templated bottom-up multiprotein nanoassembly through defined click chemistry linkage to DNA
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.
dna nanotechnology, precision assembly, origami, protein engineering, copper-free click chemistry, energy transfer, expanded genetic code
1936-0851
5003-5010
Marth, Gabriella
515790e1-1d78-4827-9cae-76947292903b
Hartley, Andrew M.
0a948101-e698-4f8f-8b56-33b74e5e5eab
Reddington, Samuel C.
e732b489-c82b-436b-ac0a-867c06b64313
Sargisson, Lauren, Lucinda
23264d2c-32e8-45c0-8083-8c0b40da08c9
Parcollet, Marlène
69da02cb-38eb-4127-9bb8-425faefd6831
Dunn, Katherine E.
bc465a08-f1b9-499a-b5a6-58dc737305ee
Jones, D. Dafydd
00ea4a6b-0a5f-4d87-8c3c-c417056211fb
Stulz, Eugen
9a6c04cf-32ca-442b-9281-bbf3d23c622d
Marth, Gabriella
515790e1-1d78-4827-9cae-76947292903b
Hartley, Andrew M.
0a948101-e698-4f8f-8b56-33b74e5e5eab
Reddington, Samuel C.
e732b489-c82b-436b-ac0a-867c06b64313
Sargisson, Lauren, Lucinda
23264d2c-32e8-45c0-8083-8c0b40da08c9
Parcollet, Marlène
69da02cb-38eb-4127-9bb8-425faefd6831
Dunn, Katherine E.
bc465a08-f1b9-499a-b5a6-58dc737305ee
Jones, D. Dafydd
00ea4a6b-0a5f-4d87-8c3c-c417056211fb
Stulz, Eugen
9a6c04cf-32ca-442b-9281-bbf3d23c622d

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

Record type: Article

Abstract

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.

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Accepted/In Press date: 17 April 2017
e-pub ahead of print date: 20 April 2017
Published date: 23 May 2017
Keywords: dna nanotechnology, precision assembly, origami, protein engineering, copper-free click chemistry, energy transfer, expanded genetic code
Organisations: CBDT, Chemistry, Molecular Assembly, Function & Structure

Identifiers

Local EPrints ID: 407989
URI: http://eprints.soton.ac.uk/id/eprint/407989
ISSN: 1936-0851
PURE UUID: ae2dbec3-fde0-4a6a-a8b7-d81405b51e73
ORCID for Eugen Stulz: ORCID iD orcid.org/0000-0002-5302-2276

Catalogue record

Date deposited: 06 May 2017 01:04
Last modified: 18 Feb 2021 17:07

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Contributors

Author: Gabriella Marth
Author: Andrew M. Hartley
Author: Samuel C. Reddington
Author: Lauren, Lucinda Sargisson
Author: Marlène Parcollet
Author: Katherine E. Dunn
Author: D. Dafydd Jones
Author: Eugen Stulz ORCID iD

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