Abstract
Introduction: The field of protein design has drastically evolved over the past four years. Both the protein folding problem, which involves predicting the 3D arrangement of atoms from a given sequence of amino acids, and its inverse, have been technically solved after 50 years. However, the black box nature of the tools developed to address these problems limits our comprehension of protein folding and dynamics. Harnessing this knowledge could revolutionise sectors such as drug design, disease diagnosis, energy transfer, and material science. This work focuses on the rational design of a protein scaffold called coiled coils, positioning them as a model for advancing our control and understanding of proteins.Results: In this thesis, we navigate the uncharted territory of coiled coils with reduced symmetry. We generate novel A3B3 hexameric α-helical barrels with both parallel and antiparallel helix orientations, expanding understanding of coiled-coil assemblies and introducing new scaffolds. Utilising these assemblies, we create covalently attached bipyridyl functional groups situated within the barrel cores, capable of chelating iron and ruthenium ions. Additionally, we develop intrinsically disordered peptide sequences that assemble only upon the introduction of specific metal ions. This can be applied for both metal sensing, as well as metal mediated sensing of other ligands.
Conclusions: This research advances the field of protein design through the generation of novel α-helical barrels and the development of coiled-coil assemblies with innovative functionalities. Our work has allowed for new potential applications in bio-sensing and catalysis and has further demonstrated the broad versatility of coiled-coil scaffolds.
Implications: This study illuminates the potential of coiled coils in the understanding of protein structure-function relationships. It introduces metal-sensitive peptide sequences for bio-sensing and photocatalysis within α-helical barrels, potentially paving the way for advancements in applications for de novo designed proteins.
| Date of Award | 5 Dec 2023 |
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| Original language | English |
| Awarding Institution |
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| Supervisor | Dek N Woolfson (Supervisor) & Alison Rodger (Supervisor) |