Identification and structural characterization of a mannose-6-phosphate containing oligomannosidic N-glycan from human erythropoietin secreted by recombinant BHK-21 cells.
A sialidase resistant mono-charged N-glycan was isolated from glycosylation site I (Asn-24) of recombinant human erythropoietin expressed from baby hamster kidney (BHK-21) cells and constituted approximately 2-4% of the oligosaccharide material at this glycosylation site. Mass spectrometry and both 1- and 2-dimensional NMR techniques revealed a high mannose type structure (Man6) with a phospho-diesterbridged N-acetylglucosamine as follows: [formula: see text]
Nimtz
M
M
Department of Molecular and Instrumental Structure Research, GBF-Gesellschaft für Biotechnologische Forschung mbH, Braunschweig, Germany.
Wray
V
V
Rüdiger
A
A
Conradt
H S
HS
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004921:Erythropoietin / Q000096:biosynthesis / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D007668:Kidney; D009682:Magnetic Resonance Spectroscopy; D008360:Mannosephosphates / Q000032:analysis*; D013058:Mass Spectrometry; D008745:Methylation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D014162:Transfection
D008360:Mannosephosphates; D009844:Oligosaccharides; D011994:Recombinant Proteins; D004921:Erythropoietin; C027693:mannose-6-phosphate