title: Synthetic Glycosphingolipids for Live-Cell Labeling.
abstract: Glycosphingolipids are an important component of cell membranes that are involved in many biological processes. Fluorescently labeled glycosphingolipids are frequently used to gain insight into their localization. However, the attachment of a fluorophore to the glycan part or-more commonly-to the lipid part of glycosphingolipids is known to alter the biophysical properties and can perturb the biological function of the probe. Presented here is the synthesis of novel glycosphingolipid probes with mono- and disaccharide head groups and ceramide moieties containing fatty acids of varying chain length (C4 to C20). These glycosphingolipids bear an azide or an alkyne group as chemical reporter to which a fluorophore can be attached through a bioorthogonal ligation reaction. The fluorescent tag and any linker connected to it can be chosen in a flexible manner. We demonstrate the suitability of the probes by selective visualization of the plasma membrane of living cells by confocal microscopy techniques. Whereas the derivatives with the shorter fatty acids can be directly applied to HEK 293T cells, the hydrophobic glycosphingolipids with longer fatty acids can be delivered to cells using fusogenic liposomes.
authors.affiliation: Department of Chemistry and ‡Department of Biology, Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Dauner
authors.affiliation: Department of Chemistry and ‡Department of Biology, Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Ellen
authors.initials: E
authors.lastname: Batroff
authors.affiliation: Department of Chemistry and ‡Department of Biology, Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: Bachmann
authors.affiliation: Department of Chemistry and ‡Department of Biology, Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Christof R
authors.initials: CR
authors.lastname: Hauck
authors.affiliation: Department of Chemistry and ‡Department of Biology, Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Valentin
authors.initials: V
authors.lastname: Wittmann
mesh_terms: D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D057930:Click Chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006028:Glycosphingolipids / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D013194:Staining and Labeling
chemical_list: D000480:Alkynes; D001386:Azides; D005456:Fluorescent Dyes; D006028:Glycosphingolipids