Title :
The carbohydrate chains of the beta
subunit of human
chorionic gonadotropin produced by the choriocarcinoma cell line BeWo
Abstract :
- Novel O-linked and novel bisecting-GlcNAc-containing N-linked carbohydrates.
- The N-linked carbohydrate chains of the beta subunit of human chorionic gonadotropin ( hCG-beta ) isolated from the culture fluid of the choriocarcinoma cell line BeWo were released enzymatically by peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase F F. Subsequently, the O-linked oligosaccharides were split off from the N-deglycosylated protein by mild alkaline borohydride treatment
- The carbohydrate chains were purified in their intact sialylated forms by FPLC anion-exchange chromatography on Mono Q, HPLC on Lichrosorb-NH2, and high-pH anion-exchange chromatography on CarboPac PA1
- 1H-NMR spectroscopic analysis of the major fractions demonstrates the occurrence of the following sialylated diantennary and triantennary N-linked oligosaccharides
- Residues not written in bold letters are variably present
- [formula: see text] The incidence of triantennary carbohydrate chains is much higher than in normal urinary hCG-beta (26% vs 2%)
- The same holds for the alpha 1-6-fucosylation of the asparagine-bound GlcNAc asparagine-bound GlcNAc (95% vs 42%)
- The presence of a bisecting GlcNAc and the occurrence of alpha 2-6-linked Neu5Ac in the most abundant N-glycans, are new features for hCG-beta
- The major O-linked carbohydrate chains identified are the tetrasaccharide Neu5Ac alpha 2-3Gal beta 1-3(Neu5Ac alpha 2-6)GalNAc-ol and the hexasaccharide Neu5Ac alpha 2-3Gal beta 1-4GlcNAc beta 1-6(Neu5Ac alpha 2-3Gal beta 1-3)GalNAc-ol, both also found in normal urinary hCG
- In addition, two novel O-glycans were characterized: [formula: see text