Selectivity and stability of N-terminal targeting protein modification chemistries

Lydia J Barber; Nicholas D J Yates; Martin A Fascione; Alison Parkin; Glyn R Hemsworth; Paul G Genever; Christopher D Spicer

doi:10.1039/d2cb00203e

Selectivity and stability of N-terminal targeting protein modification chemistries

RSC Chem Biol. 2022 Nov 17;4(1):56-64. doi: 10.1039/d2cb00203e. eCollection 2023 Jan 4.

Authors

Lydia J Barber^{1

2

3}, Nicholas D J Yates^{1

2}, Martin A Fascione^{1

2}, Alison Parkin¹, Glyn R Hemsworth⁴, Paul G Genever^{2

3}, Christopher D Spicer^{1

2

3}

Affiliations

¹ Department of Chemistry, University of York Heslington YO10 5DD UK chris.spicer@york.ac.uk.
² York Biomedical Research Institute, University of York Heslington YO10 5DD UK.
³ Department of Biology, University of York Heslington YO10 5DD UK.
⁴ Astbury Centre for Structural Molecular Biology and School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds Leeds LS2 9JT UK.

Abstract

Protein N-termini provide uniquely reactive motifs for single site protein modification. Though a number of reactions have been developed to target this site, the selectivity, generality, and stability of the conjugates formed has not been studied. We have therefore undertaken a comprehensive comparative study of the most promising methods for N-terminal protein modification, and find that there is no 'one size fits all' approach, necessitating reagent screening for a particular protein or application. Moreover, we observed limited stability in all cases, leading to a need for continued innovation and development in the bioconjugation field.