Directed assembly of defined oligomeric photosynthetic reaction centres through adaptation with programmable extra-membrane coiled-coil interfaces

Swainsbury, David J. K., Harniman, Robert L., di Bartolo, Natalie D., Liu, Juntai, Harper, William F. M., Corrie, Alexander S. and Jones, Michael R. (2016) Directed assembly of defined oligomeric photosynthetic reaction centres through adaptation with programmable extra-membrane coiled-coil interfaces. Biochimica et Biophysica Acta - Bioenergetics, 1857 (12). pp. 1829-1839. ISSN 0005-2728

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Abstract

A challenge associated with the utilisation of bioenergetic proteins in new, synthetic energy transducing systems is achieving efficient and predictable self-assembly of individual components, both natural and man-made, into a functioning macromolecular system. Despite progress with water-soluble proteins, the challenge of programming self-assembly of integral membrane proteins into non-native macromolecular architectures remains largely unexplored. In this work it is shown that the assembly of dimers, trimers or tetramers of the naturally monomeric purple bacterial reaction centre can be directed by augmentation with an α-helical peptide that self-associates into extra-membrane coiled-coil bundle. Despite this induced oligomerisation the assembled reaction centres displayed normal spectroscopic properties, implying preserved structural and functional integrity. Mixing of two reaction centres modified with mutually complementary α-helical peptides enabled the assembly of heterodimers in vitro, pointing to a generic strategy for assembling hetero-oligomeric complexes from diverse modified or synthetic components. Addition of two coiled-coil peptides per reaction centre monomer was also tolerated despite the challenge presented to the pigment-protein assembly machinery of introducing multiple self-associating sequences. These findings point to a generalised approach where oligomers or longer range assemblies of multiple light harvesting and/or redox proteins can be constructed in a manner that can be genetically-encoded, enabling the construction of new, designed bioenergetic systems in vivo or in vitro.

Item Type: Article
Additional Information: Funding Information: The authors thank Antony Burton and Dek Woolfson (School of Chemistry, University of Bristol) for help with analytical ultracentrifugation and useful discussions, and Robin Corey, Debbie Shoemark and Richard Sessions (School of Biochemistry, University of Bristol) for help and advice with molecular modelling. PeakForce atomic force microscopy was carried out in the Chemical Imaging Facility, University of Bristol with equipment funded by the Engineering and Physical Sciences Research Council (EPSRC) of the UK under grant “Atoms to Applications” (EP/K035746/1). DJKS, NDB, JL and MRJ acknowledge funding from the Biotechnology and Biological Sciences Research Council (BBSRC) of the UK (project BB/I022570/1), the BrisSynBio Synthetic Biology Research Centre (BB/L01386X/1) and the EPSRC and BBSRC Synthetic Biology Centre for Doctoral Training (EP/L016494/1). Publisher Copyright: © 2016 The Authors
Uncontrolled Keywords: coiled-coil,directed self-assembly,membrane protein,oligomerisation,photosystem,reaction centre,biophysics,biochemistry,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1304
Faculty \ School: Faculty of Science > School of Biological Sciences
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Depositing User: LivePure Connector
Date Deposited: 17 Aug 2022 12:32
Last Modified: 24 Sep 2022 07:04
URI: https://ueaeprints.uea.ac.uk/id/eprint/87372
DOI: 10.1016/j.bbabio.2016.09.002

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