Protein modification and engineering

Protein engineering

The rational engineering of proteins to generate defined functional materials is a major interest in the group. We have interests in both the application of known methodology, such as the use of bioorthogonal cross-linking chemistry and Amber suppression and the development of new methods such as the application of sortases (see below). A major current interest is the modification of the bacterial toxoids, such as that from cholera. Many members of the group are working with Mike and his close collaborator Professor Bruce Turnbull to chemically and genetically modify this class of proteins for potential application in drug delivery, the generation of protein-derived materials and sensing of unnatural glycosides.

Protein modification using Sortase

We are interested in the application of all kinds of protein modifications but have particular interests in the application of Sortases as generic tools for protein modification. We have recently reported an optimised protocol for both N-terminal and C-terminal labelling of proteins using peptide substrates. This provides a convenient, quantitative route to generate labelled proteins, superior to the use of less-selective reagents such as activated esters and readily applicable to other systems. We are now using this approach in a variety of model systems as well as seeking to develop further chemoenzymatic modification strategies.

Publications in this area

Quantitative N- or C-Terminal Protein Labelling with Unactivated Peptides by Use of Sortases and a D-Aminopeptidase Zoe L.P. Arnott, Holly E. Morgan, Kristian Hollingsworth, Charlotte M. E. Stevenson, Lawrence J. Collins, Alexandra Tamasanu, Darren C. Machin, Jonathan P. Dolan, Tomasz P. Kaminski, Gemma C. Wildsmith, Daniel J. Williamson, Isabelle B. Pickles, Stuart L. Warriner, W. Bruce Turnbull and Michael E. Webb Angew. Chem. Int Ed. (2024) 63, e202310862

Combined application of orthogonal sortases and depsipeptide substrates for dual protein labelling Holly E. Morgan, Zoe L.P. Arnott, Tomasz P. Kaminski, W. Bruce Turnbull & Michael E. Webb Bioconjugate Chem. (2022) 33 2341-2347

Synthesis of cholera toxin B-subunit glycoconjugates using site-specific orthogonal oxime and sortase ligation reactions Jonathan P Dolan, Darren C Machin, Simone Dedola, Robert A Field, Michael E Webb, W Bruce Turnbull Frontiers in Chemistry (2022) 10 958272

Challenges in the use of sortase and other peptide ligases for site-specific protein modification Holly E. Morgan, W. Bruce Turnbull and Michael E. Webb Chem. Soc. Rev. (2022) 51 4121-4145

Sortase-mediated labelling of lipid nanodiscs for cellular tracing A. Ivona Petrache, Darren C. Machin, Daniel J. Williamson, Michael E. Webb and Paul A. Beales Mol. Bio. Syst. (2016) 12 1760-1763

Confirmation of a protein-protein interaction in the pantothenate biosynthetic pathway using Sortase-mediated labelling
Philip M.Morrison, Matthew R. Balmforth, Samuel W. Ness, Daniel J. Williamson, Michael D. Rugen, W. Bruce Turnbull and Michael E. Webb ChemBioChem (2016) 17 753-758

Depsipeptide substrates for sortase-mediated N-terminal protein ligation
D. J. Williamson, M. E. Webb & W. B. Turnbull Nat. Protoc. (2014) 9 253-262

Efficient N-terminal labelling of proteins by use of Sortase
Daniel J. Williamson, Martin A. Fascione, Michael E Webb and W. Bruce Turnbull Angew. Chem. (2012) 51 9377-9380

Other protein engineering approaches

We are also interested in other approaches to modify proteins including the use of selective chemical methods to modify proteins

Publications in this area

Bio-orthogonal, bifunctional linker for engineering synthetic glycoproteins Ryan McBerney, Jonathan P Dolan, Emma E. Cawood, Michael E Webb, W Bruce Turnbull JACS Au (2022) 2, 2038–2047

Double quick, double click reversible peptide "stapling" Claire M. Grison, George M. Burslem, Jennifer A. Miles, Ludwig K.A. Pilsl, David J Yeo, Zeynab Imani, Stuart L. Warriner, Michael E. Webb, Andrew J. Wilson Chem. Sci. (2017) 8 5166-5171

Chemical generation and modification of peptides containing multiple dehydroalanines Phillip M. Morrison, Patrick J. Foley, Stuart L. Warriner & Michael E. Webb Chem. Commun. (2015) 51 13470-3