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Lookup NU author(s): Esme Hutton, Rachel Howarth, Professor Craig RobsonORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
This journal is © The Royal Society of Chemistry, 2026. The development of methods for chemical modification of proteins has enabled transformative advancements in the fields of chemical biology, cell biology and biomedicine. Herein we demonstrate that simple, easy-to-prepare acryloyl imides can be used as reagents for N-terminal protein bioconjugation that exploit conjugate addition/ring expansion (CARE) cascade reactions to afford novel protein-macrocycle conjugates. The CARE approach advantageously proceeds with high N-terminal selectivity, affords the formation of stable bioconjugates driven by irreversible ring expansion, and facilitates access to modified proteins appended with complex, functionalized medium sized rings. The utility of this late stage protein macrocycle diversification is showcased in nanobody-based imaging of cancer tissue and modulation of a chemokine–receptor interaction, uniquely decoupling GPCR endocytosis from phosphorylation. As such the CARE bioconjugation represents a powerful and versatile platform with broad potential application in the construction of tools for dissecting biological mechanisms, and biologics with new therapeutic modalities.
Author(s): Hughes OR, Tufail A, Hutton E, Nabarro J, Howarth R, Yates ND, Whitwood AC, Robson CN, Signoret N, Fascione MA, Spicer CD, Unsworth WP
Publication type: Article
Publication status: Published
Journal: Chemical Science
Year: 2026
Pages: Epub ahead of print
Online publication date: 16/12/2025
Acceptance date: 12/12/2025
Date deposited: 07/01/2026
ISSN (print): 2041-6520
ISSN (electronic): 2041-6539
Publisher: Royal Society of Chemistry
URL: https://doi.org/10.1039/d5sc08044d
DOI: 10.1039/d5sc08044d
Data Access Statement: CCDC 2391160 contains the supplementary crystallographic data for this paper (https://doi.org/10.5517/ccdc.csd.cc2l8669) The data that support the findings of this study are available in the supplementary information (SI) of this article. Supplementary information: including detailed experimental procedures for small molecule, peptide and protein transformations, compound characterization data, chromatograms, LCMS data, MALDI, kinetic methods, protein expression methods, immunofluorescence staining and microscopy and NMR spectra images. See DOI: https://doi.org/10.1039/d5sc08044d
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