Ross, JF, Wildsmith, GC, Johnson, M et al. (9 more authors) (2019) Directed Assembly of Homopentameric Cholera Toxin B‑Subunit Proteins into Higher-Order Structures Using Coiled-Coil Appendages. Journal of the American Chemical Society, 141 (13). pp. 5211-5219. ISSN 0002-7863
Abstract
The self-assembly of proteins into higher order structures is ubiquitous in living systems. It is also an essential process for the bottom-up creation of novel molecular architectures and devices for synthetic biology. However, the complexity of protein-protein interaction surfaces makes it challenging to mimic natural assembly processes in artificial systems. Indeed, many successful computationally designed protein assemblies are pre-screened for ‘designability’, limiting the choice of components. Here, we report a simple and pragmatic strategy to assemble chosen multi-subunit proteins into more complex structures. A coiled-coil domain appended to one face of the pentameric cholera toxin B-subunit (CTB) enabled the ordered assembly of tubular supra-molecular complexes. X-ray crystallography and analysis of a tubular structure has revealed a hierarchical assembly process that displays features reminiscent of the polymorphic assembly of polyomavirus proteins. The approach provides a simple and straightforward method to direct the assembly of protein building blocks which present either termini on a single face of an oligomer. This scaffolding approach can be used to generate bespoke supramolecular assemblies of functional proteins. Additionally, structural resolution of the scaffolded assemblies highlight ‘native-state’ forced protein-protein interfaces, which may prove useful as starting conformations for future computational design.
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Copyright, Publisher and Additional Information: | © 2019 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. | ||||||||||||||
Keywords: | Supramolecular assembly; protein design; cholera toxin; coiled-coil; de novo protein assembly | ||||||||||||||
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Institution: | The University of Leeds | ||||||||||||||
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) > Organic Chemistry (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cryo EM, Image Processing (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Synthetic Biology (Leed) |
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Depositing User: | Symplectic Publications | ||||||||||||||
Date Deposited: | 12 Mar 2019 13:54 | ||||||||||||||
Last Modified: | 25 Sep 2020 12:24 | ||||||||||||||
Status: | Published | ||||||||||||||
Publisher: | American Chemical Society | ||||||||||||||
Identification Number: | https://doi.org/10.1021/jacs.8b11480 | ||||||||||||||
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