The Lewis x epitope is a major non-reducing structure in the sulphated N-glycans attached to Asn-65 of bovine pro-opiomelanocortin.
The N-terminal glycopeptide of pro-opiomelanocortin (POMC), designated as the 16K fragment, is highly conserved throughout vertebrates from amphibians to mammals and is likely therefore to have an important functional role. In this paper, we report the first structural characterization of N-glycans attached to asparagine-65 of a 16K glycopeptide. The 16K fragment was isolated from bovine pituitaries and the N-glycans were analysed using fast atom bombardment mass spectrometry together with sugar and linkage analysis. Sulphated-N-acetylgalactosamine-capped antennae, typical of the pituitary glycohormones, were present in the major acidic components. The POMC oligosaccharides are distinct from those of the pituitary glycohormones because the sulphate is exclusively located on the 3-arm of biantennary structures and, in addition, a significant proportion of the molecules carry the Lewis x epitope. It is probable that these differences reflect the absence of a tripeptide motif in POMC which fully conforms to the criteria previously defined for the recognition sequence for the N-acetylgalactosamine transferase that is specific for the pituitary glycohormones [Smith and Baenziger (1992) Proc. Natl. Acad. Sci. USA, 89, 329-333]. It remains to be seen whether the Lewis x epitope is involved in selectin-mediated events, but previous studies suggest that the sulphated moieties are unlikely to play a major role in clearance. The Lewis x epitope is also present in the neutral N-linked oligosaccharides, together with a variety of other antennae including a rarely found fucosylated GalNAc-GlcNAc structure.
Siciliano
R A
RA
Department of Biochemistry, Imperial College of Science Technology and Medicine, London, UK.
Morris
H R
HR
McDowell
R A
RA
Azadi
P
P
Rogers
M E
ME
Bennett
H P
HP
Dell
A
A
D000116:Acetylgalactosamine / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D017124:Conserved Sequence; D000939:Epitopes / Q000032:analysis / Q000737:chemistry; D005643:Fucose / Q000032:analysis; D006020:Glycopeptides / Q000737:chemistry*; D007202:Indicators and Reagents; D007983:Lewis Blood Group Antigens / Q000737:chemistry*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011333:Pro-Opiomelanocortin / Q000737:chemistry* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013464:Sulfuric Acids / Q000032:analysis; D014357:Trypsin
D000939:Epitopes; D006020:Glycopeptides; D007202:Indicators and Reagents; D007983:Lewis Blood Group Antigens; D009842:Oligopeptides; D010446:Peptide Fragments; D013464:Sulfuric Acids; D005643:Fucose; D011333:Pro-Opiomelanocortin; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin; D000116:Acetylgalactosamine