title: Rapid labeling of metabolically engineered cell-surface glycoconjugates with a carbamate-linked cyclopropene reporter.
abstract: Metabolic oligosaccharide engineering is a valuable tool to monitor cellular carbohydrates. Here, we report the synthesis of a novel N-acyl-mannosamine derivative bearing a methylcyclopropene tag that is attached to the sugar via a carbamate moiety. This derivative undergoes rapid Diels-Alder reaction with inverse electron demand. We demonstrate that the cell's biosynthetic machinery incorporates this non-natural mannosamine derivative into glycoconjugates that can, subsequently, be labeled within less than 10 min with a new sulfo-Cy3-tetrazine conjugate. Using this tetrazine-dye conjugate for the detection of the methylcyclopropene-tagged mannosamine derivative, we could achieve dual labeling of two different metabolically incorporated sugars combining a Diels-Alder reaction with inverse electron demand and a strain-promoted azide-alkyne cycloaddition which are carried out simultaneously in a single step.
authors: University of Konstanz , Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), Universitätsstraße 10, 78457 Konstanz, Germany.
authors: Anne-Katrin
authors: AK
authors: Späte
authors: Holger
authors: H
authors: Bußkamp
authors: Andrea
authors: A
authors: Niederwieser
authors: Verena F
authors: VF
authors: Schart
authors: Andreas
authors: A
authors: Marx
authors: Valentin
authors: V
authors: Wittmann
mesh_terms: D002219:Carbamates / Q000737:chemistry* / Q000378:metabolism; D003521:Cyclopropanes / Q000737:chemistry* / Q000378:metabolism; D006001:Glycoconjugates / Q000737:chemistry* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D060847:Metabolic Engineering; D015394:Molecular Structure; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D013499:Surface Properties
chemical_list: D002219:Carbamates; D003521:Cyclopropanes; D006001:Glycoconjugates; D009844:Oligosaccharides; C021985:cyclopropene