Selectivity and stability of N-terminal targeting protein modification chemistries

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.

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Article

Authors/Creators:

Barber, Lydia J.
Yates, Nicholas D.J. (nicholas.yates@york.ac.uk)
Fascione, Martin A. https://orcid.org/0000-0002-0066-4419
Parkin, Alison https://orcid.org/0000-0003-4715-7200
Hemsworth, Glyn R. https://orcid.org/0000-0002-8226-1380
Genever, Paul G. https://orcid.org/0000-0002-5730-8976
Spicer, Christopher D. https://orcid.org/0000-0001-8787-578X

Funding Information: L. J. B. and C. D. S. acknowledge PhD studentship support from the BBSRC White Rose Mechanistic Biology DTP. This PhD was supported with generous industrial CASE support from Qkine and our thanks go to Dr Marko Hyvönen and Dr Catherine Elton for their support. N. D. J. Y. and M. A. F. acknowledge support from the Engineering and Physical Sciences Research Council (EPSRC, EP/V044303/1). Dr Alex Heyam is thanked for help running NMR kinetics experiments. We thank Dr Ed Bergstrom and The York Centre of Excellence in Mass Spectrometry for support with protein mass spectrometry. This centre was created thanks to a major capital investment through Science City York, supported by Yorkshire Forward with funds from the Northern Way Initiative, and subsequent support from the EPSRC (EP/K039660/1; EP/M028127/1). Publisher Copyright: © 2022 The Author(s).

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