title: High-efficiency labeling of sialylated glycoproteins on living cells.
abstract: We describe a simple method for efficiently labeling cell-surface sialic acid-containing glycans on living animal cells. The method uses mild periodate oxidation to generate an aldehyde on sialic acids, followed by aniline-catalyzed oxime ligation with a suitable tag. Aniline catalysis dramatically accelerates oxime ligation, allowing use of low concentrations of aminooxy-biotin at neutral pH to label the majority of cell-surface sialylated glycoproteins while maintaining high cell viability.
authors.affiliation: Department of Chemical Physiology, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, CA 92037, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zeng
authors.forename: T N C
authors.initials: TN
authors.lastname: Ramya
authors.forename: Anouk
authors.initials: A
authors.lastname: Dirksen
authors.forename: Philip E
authors.initials: PE
authors.lastname: Dawson
authors.forename: James C
authors.initials: JC
authors.lastname: Paulson
mesh_terms: D000447:Aldehydes / Q000737:chemistry; D000814:Aniline Compounds / Q000737:chemistry; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry; D019921:Biotinylation / Q000379:methods*; D016466:CHO Cells; D002384:Catalysis; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis; D005434:Flow Cytometry; D006801:Humans; D017493:Leukocyte Common Antigens / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D019158:N-Acetylneuraminic Acid / Q000031:analogs & derivatives / Q000737:chemistry*; D010084:Oxidation-Reduction; D010091:Oximes / Q000737:chemistry; D010504:Periodic Acid / Q000737:chemistry; D012799:Sialyltransferases / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D019809:Streptavidin / Q000737:chemistry; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
chemical_list: D000447:Aldehydes; D000814:Aniline Compounds; D008562:Membrane Glycoproteins; D010091:Oximes; D010504:Periodic Acid; D001710:Biotin; D019809:Streptavidin; C009288:metaperiodate; D012799:Sialyltransferases; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human; D019158:N-Acetylneuraminic Acid; C023650:aniline; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase