Title : Structural study of
the N-linked oligosaccharides of
hepatocyte growth factor by two-dimensional sugar mapping
Abstract :
- The structures of the N-linked oligosaccharides on recombinant human hepatocyte growth factor (rh- HGF ) expressed by Chinese hamster ovary (CHO) cells were studied by two-dimensional sugar mapping
- The oligosaccharides released from the glycopeptides by peptide: N-glycosidase F ( PNGase F) treatment were tagged with 2-aminopyridine at the reducing ends
- The alpha-chain was linked by biantennary, triantennary, and tetraantennary oligosaccharides , but the dominant oligosaccharides linking the beta-chain were biantennary (> 85%)
- There was no significant difference in oligosaccharide structures between the two glycosylation sites on each chain, that is, Asn263 and Asn371 on the alpha-chain , and Asn535 and Asn622 on the beta-chain
- The linkage of sialic acid to the non-reducing terminal galactose was identified as NeuAc alpha(2-3) by 1H-NMR spectrometry
- The structures of the N-linked oligosaccharides from rat HGF were also studied
- Triantennary oligosaccharides were obtained from the alpha-chain and a biantennary oligosaccharide was obtained from the beta-chain
- This result indicates that the alpha-chain is also linked by higher branched oligosaccharides than the beta-chain in rat HGF