Title

An intrinsic membrane glycoprotein of the golgi apparatus with O-linked N-acetylglucosamine facing the cytosol

UMMS Affiliation

Department of Biochemistry

Date

12-25-1988

Document Type

Article

Subjects

Acetylglucosamine; Animals; Cell Fractionation; Cytosol; Endoplasmic Reticulum; Glucosamine; Golgi Apparatus; Intracellular Membranes; Liver; Male; Membrane Glycoproteins; Molecular Weight; Oligosaccharides; Rats; Rats, Inbred Strains

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

We have recently described the occurrence of integral membrane glycoproteins in rat liver smooth and rough endoplasmic reticulum with O-N-acetylglucosamine facing the cytosolic and luminal sides of the membrane (Abeijon, C., and Hirschberg, C. B. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 1010-1014). We now report that integral membrane glycoproteins with cytosolic facing O-N-acetylglucosamine also occur in membranes of rat liver Golgi apparatus. This was determined following incubation of vesicles from the Golgi apparatus, which were sealed and of the same membrane topographical orientation as in vivo, with UDP-[14C]galactose and saturating amounts of bovine milk galactosyltransferase. This enzyme does not enter the lumen of the vesicles and specifically catalyzes the addition of galactose, in a beta 1-4 linkage, to terminal N-acetylglucosamine. Under these conditions, galactose was transferred to a glycoprotein of molecular mass of 92 kDa. This protein was insoluble in sodium carbonate, pH 11.5, conditions under which integral membrane proteins remain membrane bound and was insensitive to treatment with peptide:N-glycosidase F. beta Elimination and chromatography showed that radiolabeled galactose was part of a disaccharide which was characterized as Gal beta 1-4GlcNAcitol. This glycoprotein is specific of the Golgi apparatus membrane. Intrinsic membrane glycoproteins with this unusual carbohydrate membrane orientation thus occur in the endoplasmic reticulum and Golgi apparatus of rat liver.

Rights and Permissions

Citation: J Biol Chem. 1988 Dec 25;263(36):19778-82.

Related Resources

Link to Article in PubMed

Journal Title

The Journal of biological chemistry

PubMed ID

3198650