The glycosylation of Bowes melanoma tissue plasminogen activator: lectin mapping, reaction with anti-L2/HNK-1 antibodies and the presence of sulphated/glucuronic acid containing glycans.
The glycosylation of tissue plasminogen activator (t-PA) obtained from the Bowes melanoma cell line was re-examined using methods of serial lectin affinity chromatography coupled with Bio-Gel P-4 gel filtration chromatography and exoglycosidase sequencing. This study clarified an earlier discrepancy in the literature and confirmed that the major complex N-linked glycans on Bowes t-PA that carry sialic acid as their sole charged group are bi-antennary, core fucosylated, with terminal N-acetylgalactosamine residues. We also report the characterization of a series of related and previously unidentified sialylated glycans. Further we show that Bowes t-PA expresses glucuronic acid/sulphate containing N-linked glycans and is recognized by anti-carbohydrate L2/HNK-1 monoclonal antibodies. The presence on Bowes t-PA of glycans associated primarily with the nervous system is consistent with its expression in a cell line of neuroectodermal origin.
Jaques
A J
AJ
Biochemistry Department, Oxford University, U.K.
Opdenakker
G
G
Rademacher
T W
TW
Dwek
R A
RA
Zamze
S E
SE
D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D018959:CD57 Antigens / Q000276:immunology; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004590:Electrophoresis, Paper; D000939:Epitopes / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D005965:Glucuronates / Q000032:analysis; D006031:Glycosylation; D037102:Lectins / Q000378:metabolism; D008546:Melanoma, Experimental / Q000737:chemistry*; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis*; D012794:Sialic Acids / Q000378:metabolism; D013431:Sulfates / Q000032:analysis; D010959:Tissue Plasminogen Activator / Q000737:chemistry* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D018959:CD57 Antigens; D000939:Epitopes; D005965:Glucuronates; D037102:Lectins; D009844:Oligosaccharides; D011134:Polysaccharides; D012794:Sialic Acids; D013431:Sulfates; C073031:glucuronic acid 3-sulfate; D010959:Tissue Plasminogen Activator; D019158:N-Acetylneuraminic Acid