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. 1990 Dec;87(24):9789–9793. doi: 10.1073/pnas.87.24.9789

An intrinsic membrane glycoprotein with cytosolically oriented n-linked sugars.

C H Pedemonte 1, G Sachs 1, J H Kaplan 1
PMCID: PMC55259  PMID: 2175915

Abstract

We demonstrate that the Na(+)-pump alpha-subunit polypeptide is glycosylated by using bovine milk galactosyltransferase, a specific enzyme which attaches galactose to terminal N-acetylglucosamine residues. The galactose acceptor sites are available for glycosylation only after permeabilization of right-side-out vesicles prepared from kidney outer medulla; therefore, the oligosaccharide moieties are facing the cytoplasm of the cell. We further show that the oligosaccharides are bound to asparagine residues of the alpha-subunit polypeptide, since the protein-carbohydrate linkage is hydrolyzed by peptide-N glycosidase F (an enzyme specific for N-linked sugars). Thus, the Na(+)-pump alpha subunit is a glycoprotein with its N-linked oligosaccharide moieties located at the cytosolic face of the cell membrane. Intrinsic membrane glycoproteins with such an oligosaccharide-protein linkage and cell membrane orientation have not been previously reported, to our knowledge.

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Selected References

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