Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry

Worthy, Harley L., Sabah Auhim, Husam, Jamieson, W. David ORCID: https://orcid.org/0000-0001-8260-5211, Pope, Jacob R. ORCID: https://orcid.org/0000-0001-6478-5378, Wall, Aaron, Batchlor, Robert, Johnson, Rachel L., Watkins, Daniel W. ORCID: https://orcid.org/0000-0003-3825-5036, Rizkallah, Pierre ORCID: https://orcid.org/0000-0002-9290-0369, Castell, Oliver K. ORCID: https://orcid.org/0000-0002-6059-8062 and Jones, D. Dafydd ORCID: https://orcid.org/0000-0001-7709-3995 2019. Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry. Communications Chemistry 2 , 83. 10.1038/s42004-019-0185-5

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Abstract

Construction of artificial higher order protein complexes allows sampling of structural architectures and functional features not accessible by classical monomeric proteins. Here, we combine in silico modelling with expanded genetic code facilitated strain promoted azide-alkyne cycloaddition to construct artificial complexes that are structurally integrated protein dimers and demonstrate functional synergy. Using fluorescent proteins sfGFP and Venus as models, homodimers and heterodimers are constructed that switched ON once assembled and display enhanced spectral properties. Symmetrical crosslinks are found to be important for functional enhancement. The determined molecular structure of one artificial dimer shows that a new long-range polar network comprised mostly of organised water molecules links the two chromophores leading to activation and functional enhancement. Single molecule analysis reveals the dimer is more resistant to photobleaching spending longer times in the ON state. Thus, genetically encoded bioorthogonal chemistry can be used to generate truly integrated artificial protein complexes that enhance function.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Biosciences Pharmacy Advanced Research Computing @ Cardiff (ARCCA)
Additional Information:	This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.
Publisher:	Nature Research
ISSN:	2399-3669
Date of First Compliant Deposit:	8 July 2019
Date of Acceptance:	21 June 2019
Last Modified:	11 Oct 2023 20:53
URI:	https://orca.cardiff.ac.uk/id/eprint/124076

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