Kinetics and stability of N-Terminal Protein modification strategies L. J. Barber* 1,2,3 , P. Genever 2,3 , C. D. Spicer 1,3 1 Department of Chemistry, University of York, UK, 2 York Biomedical Research Institute, University of York, UK, 3 Department of Biology, University of York, UK N-terminal targeting has emerged as a powerful means to functionalise proteins, for example in the synthesis of protein-polymer conjugates for tissue engineering or antibody drug conjugates. 1 However, selectivity is often poor or the conjugates formed suffer from instability; 2 identifying the most suitable N-terminal modification strategy for an intended application is therefore critical. We have undertaken a detailed comparative study of the conversion, selectivity, and stability of leading N-terminal modification strategies to provide key insight into the formation and utility of the resultant protein-conjugates. 3 Critically, all N-terminal modification strategies were found to exhibit slow kinetics and some extent of reversibility, with reaction efficiency and selectivity found to be highly protein dependent: there is no “one size fits all” approach to N-terminal protein modification. This work highlights the need for the screening of a toolbox of complementary N-terminal modification strategies to ensure optimal properties are achieved for a given target protein and application. We are now using this work as a platform to develop new modification strategies that address current limitations, including the use of proximity-driven chemistries, and intramolecular cyclisation/hydrogen bonding.
References 1. M. B. Francis et al. Nat. Chem. Biol. 13: 697-705, 2017. 2. M. B. Francis et al. Nat. Chem. Biol. 11: 326-331, 2015. 3. C. D. Spicer et al. RSC Chem. Biol. 4: 56-64, 2023.
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© The Author(s), 2023
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