Post-translational addition of chondroitin sulfate glycosaminoglycans. Role of N-linked oligosaccharide addition, trimming, and processing.
A melanoma proteoglycan model system has been used to examine the role of core protein asparagine-linked (N-linked) oligosaccharides in the transport and assembly of proteoglycan molecules. The use of agents which block discrete steps in the trimming and processing of core oligosaccharides (castanospermine, 1-deoxynojirimycin, N-methyldeoxynojirimycin, 1-deoxymannojirimycin, and swainsonine) demonstrates that removal of glucose residues from the N-linked oligosaccharides is required for the cell surface expression of a melanoma proteoglycan core protein and for the conversion of the core protein to a chondroitin sulfate proteoglycan. However, complete maturation of the oligosaccharides to a "complex" form is not required for these events. Treatment of M21 human melanoma cells with the glucosidase inhibitors castanospermine, 1-deoxynojirimycin, or N-methyldeoxynojirimycin results in a dose-dependent inhibition of glycosaminoglycan (GAG) addition to the melanoma antigen recognized by monoclonal antibody 9.2.27. In contrast, treatment with the mannosidase inhibitors 1-deoxymannojirimycin and swainsonine does not effect GAG addition. Identical results are obtained when the major histocompatibility complex class II antigen gamma chain proteoglycan is examined in inhibitor-treated melanoma and B-lymphoblastoid cells. These data, in conjunction with the known effects of the glucosidase and mannosidase inhibitors on the transport and secretion of other glycoproteins support the hypothesis that the addition, trimming, and processing of N-linked oligosaccharides is involved in the transport of certain proteoglycan core proteins to the site of GAG addition and to the cell surface.
Spiro
R C
RC
Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
Casteel
H E
HE
Laufer
D M
DM
Reisfeld
R A
RA
Harper
J R
JR
D000951:Antigens, Neoplasm; D002460:Cell Line; D002807:Chondroitin / Q000031:analogs & derivatives*; D002809:Chondroitin Sulfates / Q000378:metabolism*; D005959:Glucosidases / Q000037:antagonists & inhibitors; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D008361:Mannosidases / Q000037:antagonists & inhibitors; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D009844:Oligosaccharides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014415:Tunicamycin / Q000494:pharmacology
D000951:Antigens, Neoplasm; D000949:Histocompatibility Antigens Class II; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D009844:Oligosaccharides; D014415:Tunicamycin; D002807:Chondroitin; D002809:Chondroitin Sulfates; D005959:Glucosidases; D008361:Mannosidases