title: Terminal alkenes as versatile chemical reporter groups for metabolic oligosaccharide engineering.
abstract: The Diels-Alder reaction with inverse electron demand (DAinv reaction) of 1,2,4,5-tetrazines with electron rich or strained alkenes was proven to be a bioorthogonal ligation reaction that proceeds fast and with high yields. An important application of the DAinv reaction is metabolic oligosaccharide engineering (MOE) which allows the visualization of glycoconjugates in living cells. In this approach, a sugar derivative bearing a chemical reporter group is metabolically incorporated into cellular glycoconjugates and subsequently derivatized with a probe by means of a bioorthogonal ligation reaction. Here, we investigated a series of new mannosamine and glucosamine derivatives with carbamate-linked side chains of varying length terminated by alkene groups and their suitability for labeling cell-surface glycans. Kinetic investigations showed that the reactivity of the alkenes in DAinv reactions increases with growing chain length. When applied to MOE, one of the compounds, peracetylated N-butenyloxycarbonylmannosamine, was especially well suited for labeling cell-surface glycans. Obviously, the length of its side chain represents the optimal balance between incorporation efficiency and speed of the labeling reaction. Sialidase treatment of the cells before the bioorthogonal labeling reaction showed that this sugar derivative is attached to the glycans in form of the corresponding sialic acid derivative and not epimerized to another hexosamine derivative to a considerable extent.
authors.affiliation: University of Konstanz, Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), 78457 Konstanz (Germany), Fax: (+49) 7531-88-4573.
authors.forename: Anne-Katrin
authors.initials: AK
authors.lastname: Späte
authors.forename: Verena F
authors.initials: VF
authors.lastname: Schart
authors.forename: Sophie
authors.initials: S
authors.lastname: Schöllkopf
authors.forename: Andrea
authors.initials: A
authors.lastname: Niederwieser
authors.forename: Valentin
authors.initials: V
authors.lastname: Wittmann
mesh_terms: D000475:Alkenes / Q000737:chemistry*; D057930:Click Chemistry; D061565:Cycloaddition Reaction; D005944:Glucosamine / Q000031:analogs & derivatives*; D057809:HEK293 Cells; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry; D006595:Hexosamines / Q000737:chemistry*; D006801:Humans; D009844:Oligosaccharides / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis*
chemical_list: D000475:Alkenes; D006573:Heterocyclic Compounds, 1-Ring; D006595:Hexosamines; D009844:Oligosaccharides; D011134:Polysaccharides; C008322:mannosamine; D005944:Glucosamine
keywords: bioorthogonal chemistry; carbohydrates; cycloaddition; metabolic oligosaccharide engineering; tetrazines