Identification and structural characterization of a mannose‐6‐phosphate containing oligomannosidic N‐glycan from human erythropoietin secreted by recombinant BHK‐21 cells

FEBS Letters

Rüdiger, Angelika¹  Nimtz, Manfred¹  Wray, Victor¹  Conradt, Harald S.² 

[1] Department of Molecular and Instrumental Structure Research, GBF-Gesellschaft für Biotechnologische Forschung mbH, Mascheroder Weg 1, D-38124 Braunschweig, Germany;Department of Cell Biology and Genetics, GBF- Gesellschaft fur Biotechnologische Forschung mbH, Mascheroder Weg 1, D-38124 Braunschweig, Germany

关键词: Recombinant human erythropoietin;    Baby hamster kidney cells (BHK-21);    Phosporylated oligomannosidic glycan;    Mannose-6-phosphate;    1- and 2-D NMR spectroscopy;    Lysosomal recognition signal;    BHK;    baby hamster kidney;    CHO;    Chinese hamster ovary;    COSY;    correlation spectroscopy;    EPO;    erythropoietin;    GC;    gas chromatography;    GlcNAc;    N-acetylglucosamine;    Hex;    hexose;    HexNAc;    N-acetylhexosamine;    HPAE-PAD;    high-pH anion-exchange chromatography with pulsed amperometric detection;    HPLC;    high performance liquid chromatography;    MALDI/TOF MS;    matrix-assisted laser desorption/ionization time-of-flight mass spectrometry;    Man;    mannose;    NMR;    nuclear magnetic resonance;    NOESY;    nuclear Overhauser enhancement and exchange spectroscopy;    PNGase F;    polypeptidase: N 4-(N-acetyl-glucosaminyl)asp aragine-amidase F;    TOCSY;    total correlation spectroscopy;   

DOI  :  10.1016/0014-5793(95)00473-M

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A sialidase resistant mono-charged N-glycan was isolated from glycosylation site I (Asn-24) of recombinant human eruthropoietin 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:

【 授权许可】

Unknown