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Directed assembly of homo-pentameric cholera toxin B-subunit proteins into higher-order structures using coiled-coil appendages

DOI: 10.1021/jacs.8b11480 DOI Help

Authors: James F Ross (University of Leeds) , Gemma C. Wildsmith (University of Leeds) , Michael Johnson (University of Leeds) , Daniel Hurdiss (University of Leeds) , Kristian Hollingsworth (University of Leeds) , Rebecca F. Thompson (University of Leeds) , Majid Mosayebi (University of Bristol) , Chi H. Trinh (University of Leeds) , Emanuele Paci (University of Leeds) , Arwen R. Pearson (Universitat Hamburg; University of Leeds) , Michael E. Webb (University of Leeds) , W. Bruce Turnbull (University of Leeds)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of The American Chemical Society

State: Published (Approved)
Published: March 2019

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.

Journal Keywords: supramolecular assembly; protein design; cholera toxin; coiled-coil; de novo proetin assembly

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I24-Microfocus Macromolecular Crystallography