Synthesis of cholera toxin B subunit glycoconjugates using site-specific orthogonal oxime and sortase ligation reactions

P. Dolan, Jonathan; Machin, Darren C.; Dedola, Simone; Field2, Robert A.; Webb, Michael E.; Bruce Turnbull, W.

doi:10.3389/fchem.2022.958272

Synthesis of cholera toxin B subunit glycoconjugates using site-specific orthogonal oxime and sortase ligation reactions

P. Dolan, Jonathan; Machin, Darren C.; Dedola, Simone; Field2, Robert A.; Webb, Michael E.; Bruce Turnbull, W.

Authors

Jonathan Dolan j.dolan@keele.ac.uk

Darren C. Machin

Simone Dedola

Robert A. Field2

Michael E. Webb

W. Bruce Turnbull

Abstract

<jats:p>The chemoenzymatic synthesis of a series of dual N- and C-terminal–functionalized cholera toxin B subunit (CTB) glycoconjugates is described. Mucin 1 peptides bearing different levels of Tn antigen glycosylation [MUC1(Tn)] were prepared <jats:italic>via</jats:italic> solid-phase peptide synthesis. Using sortase-mediated ligation, the MUC1(Tn) epitopes were conjugated to the C-terminus of CTB in a well-defined manner allowing for high-density display of the MUC1(Tn) epitopes. This work explores the challenges of using sortase-mediated ligation in combination with glycopeptides and the practical considerations to obtain high levels of conjugation. Furthermore, we describe methods to combine two orthogonal labeling methodologies, oxime- and sortase-mediated ligation, to expand the biochemical toolkit and produce dual N- and C-terminal–labeled conjugates.</jats:p>

Journal Article Type	Article
Acceptance Date	Jul 26, 2022
Publication Date	Sep 14, 2022
Journal	Frontiers in chemistry
Publisher	Frontiers Media
DOI	https://doi.org/10.3389/fchem.2022.958272
Publisher URL	https://www.frontiersin.org/articles/10.3389/fchem.2022.958272/full
PMID	36186584