title: Expanding the scope of cyclopropene reporters for the detection of metabolically engineered glycoproteins by Diels-Alder reactions.
abstract: Monitoring glycoconjugates has been tremendously facilitated by the development of metabolic oligosaccharide engineering. Recently, the inverse-electron-demand Diels-Alder reaction between methylcyclopropene tags and tetrazines has become a popular ligation reaction due to the small size and high reactivity of cyclopropene tags. Attaching the cyclopropene tag to mannosamine via a carbamate linkage has made the reaction even more efficient. Here, we expand the application of cyclopropene tags to N-acylgalactosamine and N-acylglucosamine derivatives enabling the visualization of mucin-type O-glycoproteins and O-GlcNAcylated proteins through Diels-Alder chemistry. Whereas the previously reported cyclopropene-labeled N-acylmannosamine derivative leads to significantly higher fluorescence staining of cell-surface glycoconjugates, the glucosamine derivative gave higher labeling efficiency with protein preparations containing also intracellular proteins.
authors.affiliation: University of Konstanz, Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), Universitätsstraße 10, 78457 Konstanz, Germany.
authors.forename: Anne-Katrin
authors.initials: AK
authors.lastname: Späte
authors.affiliation: University of Konstanz, Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), Universitätsstraße 10, 78457 Konstanz, Germany.
authors.forename: Verena F
authors.initials: VF
authors.lastname: Schart
authors.affiliation: University of Konstanz, Department of Biology and Konstanz Research School Chemical Biology (KoRS-CB), Universitätsstraße 10, 78457 Konstanz, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Häfner
authors.affiliation: University of Konstanz, Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), Universitätsstraße 10, 78457 Konstanz, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Niederwieser
authors.affiliation: University of Konstanz, Department of Biology and Konstanz Research School Chemical Biology (KoRS-CB), Universitätsstraße 10, 78457 Konstanz, Germany.
authors.forename: Thomas U
authors.initials: TU
authors.lastname: Mayer
authors.affiliation: University of Konstanz, Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), Universitätsstraße 10, 78457 Konstanz, Germany.
authors.forename: Valentin
authors.initials: V
authors.lastname: Wittmann
keywords: bioorthogonal chemistry; carbohydrates; cyclopropenes; inverse-electron-demand Diels-Alder reactions; metabolic oligosaccharide engineering