Metabolic labeling of glycans with azido sugars and subsequent glycan-profiling and visualization via Staudinger ligation
作者： Scott T Laughlin , Carolyn R Bertozzi
关键词: Azide 、 Bioorthogonal chemical reporter 、 Glycoprotein 、 Biophysics 、 Proteomics 、 Glycomics 、 Glycoconjugate 、 Biology 、 Glycan 、 Biochemistry 、 Drug design
摘要: Metabolic labeling of glycans with a bioorthogonal chemical reporter such as the azide enables their visualization in cells and organisms well enrichment specific glycoprotein types for proteomic analysis. This process involves two steps. Azido sugars are fed to or integrated by glycan biosynthetic machinery into various glycoconjugates. The azido then covalently tagged imaging probes epitope tags, either ex vivo vivo, using an azide-specific reaction. protocol details syntheses N-azidoacetylmannosamine (ManNAz), N-azidoacetylgalactosamine (GalNAz), N-azidoacetylglucosamine (GlcNAz) 6-azidofucose (6AzFuc), detection reagents phosphine-FLAG phosphine-FLAG-His6. Applications cellular glycoproteins analysis described. synthesis (ManNAz, GalNAz, GlcNAz 6AzFuc) (phosphine-FLAG phosphine-FLAG-His6) can be completed approximately 1 week. A cell metabolic experiment 4 d.
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