Publications by category
Journal articles
Pope JR, Johnson RL, Jamieson WD, Worthy HL, Kailasam S, Ahmed RD, Taban I, Auhim HS, Watkins DW, Rizkallah PJ, et al (2020). Association of Fluorescent Protein Pairs and its Significant Impact on Fluorescence and Energy Transfer. Advanced Science, 8(1), 2003167-2003167.
Worthy HL, Auhim HS, Jamieson WD, Pope JR, Wall A, Batchelor R, Johnson RL, Watkins DW, Rizkallah P, Castell OK, et al (2019). Author Correction: Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry (Communications Chemistry, (2019), 2, 1, (83), 10.1038/s42004-019-0185-5).
Communications Chemistry,
2(1).
Abstract:
Author Correction: Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry (Communications Chemistry, (2019), 2, 1, (83), 10.1038/s42004-019-0185-5)
© 2019, the Author(s). An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Abstract.
Gwyther REA, Dafydd Jones D, Worthy HL (2019). Better together: Building protein oligomers naturally and by design.
Biochemical Society Transactions,
47(6), 1773-1780.
Abstract:
Better together: Building protein oligomers naturally and by design
© 2019 the Author(s). Protein oligomers are more common in nature than monomers, with dimers being the most prevalent final structural state observed in known structures. From a biological perspective, this makes sense as it conserves vital molecular resources that may be wasted simply by generating larger single polypeptide units, and allows new features such as cooperativity to emerge. Taking inspiration from nature, protein designers and engineers are now building artificial oligomeric complexes using a variety of approaches to generate new and useful supramolecular protein structures. Oligomerisation is thus offering a new approach to sample structure and function space not accessible through simply tinkering with monomeric proteins.
Abstract.
Full text.
Worthy HL, Auhim HS, Jamieson WD, Pope JR, Wall A, Batchelor R, Johnson RL, Watkins DW, Rizkallah P, Castell OK, et al (2019). Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry.
Communications Chemistry,
2(1).
Abstract:
Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry
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.
Abstract.
Author URL.
Thomas SK, Jamieson WD, Gwyther REA, Bowen BJ, Beachey A, Worthy HL, Macdonald JE, Elliott M, Castell OK, Jones DD, et al (2019). Site-Specific Protein Photochemical Covalent Attachment to Carbon Nanotube Side Walls and its Electronic Impact on Single Molecule Function.
Bioconjugate Chemistry,
31(3), 584-594.
Full text.
Freeley M, Worthy HL, Ahmed R, Bowen B, Watkins D, Macdonald JE, Zheng M, Jones DD, Palma M (2017). Site-Specific One-to-One Click Coupling of Single Proteins to Individual Carbon Nanotubes: a Single-Molecule Approach. Journal of the American Chemical Society, 139(49), 17834-17840.
Halliwell LM, Jathoul AP, Bate JP, Worthy HL, Anderson JC, Jones DD, Murray JAH (2017). ΔFlucs: Brighter Photinus pyralis. firefly luciferases identified by surveying consecutive single amino acid deletion mutations in a thermostable variant. Biotechnology and Bioengineering, 115(1), 50-59.
Hartley AM, Worthy HL, Reddington SC, Rizkallah PJ, Jones DD (2016). Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle.
Chem Sci,
7(10), 6484-6491.
Abstract:
Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle.
Through the genetic incorporation of a single phenyl azide group into superfolder GFP (sfGFP) at residue 148 we provide a molecular description of how this highly versatile chemical handle can be used to positively switch protein function in vitro and in vivo via either photochemistry or bioconjugation. Replacement of H148 with p-azido-l-phenylalanine (azF) blue shifts the major excitation peak ∼90 nm by disrupting the H-bond and proton transfer network that defines the chromophore charged state. Bioorthogonal click modification with a simple dibenzylcyclooctyne or UV irradiation shifts the neutral-anionic chromophore equilibrium, switching fluorescence to the optimal ∼490 nm excitation. Click modification also improved quantum yield over both the unmodified and original protein. Crystal structures of both the click modified and photochemically converted forms show that functional switching is due to local conformational changes that optimise the interaction networks surrounding the chromophore. Crystal structure and mass spectrometry studies of the irradiated protein suggest that the phenyl azide converts to a dehydroazepine and/or an azepinone. Thus, protein embedded phenyl azides can be used beyond simple photocrosslinkers and passive conjugation handles, and mimic many natural post-translational modifications: modulation though changes in interaction networks.
Abstract.
Author URL.
Publications by year
2020
Pope JR, Johnson RL, Jamieson WD, Worthy HL, Kailasam S, Ahmed RD, Taban I, Auhim HS, Watkins DW, Rizkallah PJ, et al (2020). Association of Fluorescent Protein Pairs and its Significant Impact on Fluorescence and Energy Transfer. Advanced Science, 8(1), 2003167-2003167.
2019
Worthy HL, Auhim HS, Jamieson WD, Pope JR, Wall A, Batchelor R, Johnson RL, Watkins DW, Rizkallah P, Castell OK, et al (2019). Author Correction: Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry (Communications Chemistry, (2019), 2, 1, (83), 10.1038/s42004-019-0185-5).
Communications Chemistry,
2(1).
Abstract:
Author Correction: Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry (Communications Chemistry, (2019), 2, 1, (83), 10.1038/s42004-019-0185-5)
© 2019, the Author(s). An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Abstract.
Gwyther REA, Dafydd Jones D, Worthy HL (2019). Better together: Building protein oligomers naturally and by design.
Biochemical Society Transactions,
47(6), 1773-1780.
Abstract:
Better together: Building protein oligomers naturally and by design
© 2019 the Author(s). Protein oligomers are more common in nature than monomers, with dimers being the most prevalent final structural state observed in known structures. From a biological perspective, this makes sense as it conserves vital molecular resources that may be wasted simply by generating larger single polypeptide units, and allows new features such as cooperativity to emerge. Taking inspiration from nature, protein designers and engineers are now building artificial oligomeric complexes using a variety of approaches to generate new and useful supramolecular protein structures. Oligomerisation is thus offering a new approach to sample structure and function space not accessible through simply tinkering with monomeric proteins.
Abstract.
Full text.
Worthy HL, Auhim HS, Jamieson WD, Pope JR, Wall A, Batchelor R, Johnson RL, Watkins DW, Rizkallah P, Castell OK, et al (2019). Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry.
Communications Chemistry,
2(1).
Abstract:
Positive functional synergy of structurally integrated artificial protein dimers assembled by Click chemistry
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.
Abstract.
Author URL.
Thomas SK, Jamieson WD, Gwyther REA, Bowen BJ, Beachey A, Worthy HL, Macdonald JE, Elliott M, Castell OK, Jones DD, et al (2019). Site-Specific Protein Photochemical Covalent Attachment to Carbon Nanotube Side Walls and its Electronic Impact on Single Molecule Function.
Bioconjugate Chemistry,
31(3), 584-594.
Full text.
2017
Freeley M, Worthy HL, Ahmed R, Bowen B, Watkins D, Macdonald JE, Zheng M, Jones DD, Palma M (2017). Site-Specific One-to-One Click Coupling of Single Proteins to Individual Carbon Nanotubes: a Single-Molecule Approach. Journal of the American Chemical Society, 139(49), 17834-17840.
Halliwell LM, Jathoul AP, Bate JP, Worthy HL, Anderson JC, Jones DD, Murray JAH (2017). ΔFlucs: Brighter Photinus pyralis. firefly luciferases identified by surveying consecutive single amino acid deletion mutations in a thermostable variant. Biotechnology and Bioengineering, 115(1), 50-59.
2016
Hartley AM, Worthy HL, Reddington SC, Rizkallah PJ, Jones DD (2016). Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle.
Chem Sci,
7(10), 6484-6491.
Abstract:
Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle.
Through the genetic incorporation of a single phenyl azide group into superfolder GFP (sfGFP) at residue 148 we provide a molecular description of how this highly versatile chemical handle can be used to positively switch protein function in vitro and in vivo via either photochemistry or bioconjugation. Replacement of H148 with p-azido-l-phenylalanine (azF) blue shifts the major excitation peak ∼90 nm by disrupting the H-bond and proton transfer network that defines the chromophore charged state. Bioorthogonal click modification with a simple dibenzylcyclooctyne or UV irradiation shifts the neutral-anionic chromophore equilibrium, switching fluorescence to the optimal ∼490 nm excitation. Click modification also improved quantum yield over both the unmodified and original protein. Crystal structures of both the click modified and photochemically converted forms show that functional switching is due to local conformational changes that optimise the interaction networks surrounding the chromophore. Crystal structure and mass spectrometry studies of the irradiated protein suggest that the phenyl azide converts to a dehydroazepine and/or an azepinone. Thus, protein embedded phenyl azides can be used beyond simple photocrosslinkers and passive conjugation handles, and mimic many natural post-translational modifications: modulation though changes in interaction networks.
Abstract.
Author URL.