Apical transport of osteopontin is independent of N-glycosylation and sialylation.
Studies of how epithelial surface polarity into apical and basolateral domains is generated and maintained have proposed that carbohydrate modifications serve as apical targeting signals for proteins by interacting with lectin sorters. However, the experimental evidence in support of N-glycans, O-glycans and sialic acids mediating apical transport is still very controversial. This could be partly due to the fact that in most studies exogenously expressed proteins were analysed. One has, therefore, examined the role of carbohydrate moieties in apical targeting of the endogenous secretory protein osteopontin in MDCK cells. It was found, however, that sorting of osteopontin does not require N-glycosylation of the protein itself nor that of other factors involved in the sorting process. Incubation of cells with the inhibitor of O-glycosylation benzyl-alpha-GaINAc reduced the molecular weight of osteopontin by blocking sialic acid addition to O-glycans. Interestingly, also impairment of sialylation had no effect on polar secretion of the protein. Thus, the results show that both N-glycans and sialic acids are not essential sorting signals, suggesting that inner core carbohydrates and/or a proteinaceous signal mediate apical targeting of osteopontin.
Trischler
M
M
Institut für Biochemie, Universität Mainz, Germany.
Koch-Brandt
C
C
Ullrich
O
O
D000116:Acetylgalactosamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D001593:Benzyl Compounds / Q000494:pharmacology; D002460:Cell Line; D016764:Cell Polarity; D004285:Dogs; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006031:Glycosylation / Q000187:drug effects; D007668:Kidney / Q000166:cytology; D037102:Lectins / Q000378:metabolism; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism; D053495:Osteopontin; D011487:Protein Conformation; D021382:Protein Sorting Signals; D021381:Protein Transport; D012794:Sialic Acids / Q000378:metabolism; D012795:Sialoglycoproteins / Q000378:metabolism*
D001593:Benzyl Compounds; D037102:Lectins; D009844:Oligosaccharides; D021382:Protein Sorting Signals; D012794:Sialic Acids; D012795:Sialoglycoproteins; D053495:Osteopontin; C080739:benzyl-alpha-N-acetylgalactosamine; D000116:Acetylgalactosamine