Professor Mike Webb

Talk: Harnessing transpeptidases to engineer biomolecule function

Controlled engineering of proteins is a critical technique for both biophysics and biotechnology. We have recently described an optimised approach for quantitative labelling of proteins at either the N- or C-terminus using the combined action of transpeptidases and aminopeptidases [1]. In this approach, the transpeptidase chemoselectively modifies a protein substrate in a reversible fashion while the aminopeptidase drives the reaction to completion by selectively hydrolysing the reaction by-product. This enables quantitative protein labelling at a 1:1 protein-to-labelling peptide ratio, essentially. We have further characterised this system and will show how we can effectively label proteins expressed with common bacterial expression vectors without additional genetic modification.

In this talk, I will also describe how we have exploited this enzyme reaction using contemporary optimisation approaches to produce proteins for biophysical assays with little or any need for clean-up steps and to generate conjugated biopharmaceuticals.

[1] 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