PMID: 6411739Sep 1, 1983Paper

Transport and topology of galactosyltransferase in endomembranes of HeLa cells

The Journal of Cell Biology
G J StrousE G Berger

Abstract

HeLa cell membranes were studied for the distribution and orientation of the Golgi marker enzyme uridine diphosphate-galactose:beta-D-N-acetylglucosamine beta, 1-4 transferase (GT). Short pulse labeling in the presence of [35S]methionine resulted in two precursor species (Mr = 44,000 and 47,000), present in a microsomal fraction with a density of 1.18 g/ml in sucrose, presumably derived from the rough endoplasmic reticulum. Processing of the N-linked oligosaccharide(s) occurred only after the precursor molecules migrated to lighter density fractions, presumably derived from the Golgi complex. The mature GT molecules (Mr = 54,000) contain O-linked oligosaccharides as shown by beta-elimination of metabolically incorporated [3H]galactose. The O-glycosylation occurred mainly in the light density fractions. The topology of GT was studied on membrane fractions after labeling with [35S]methionine as well as immunocytochemically on ultrathin cryosections at the electron microscope level. Our results indicate that both the antigenic determinants of GT as well as polypeptide chain are present intramembraneously and at the luminal side of the membranes of the Golgi complex and rough endoplasmic reticulum.

References

Nov 20, 1969·Biochemical and Biophysical Research Communications·J MorreT W Keenan
Mar 1, 1981·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·E G BergerU Schilt
Jan 1, 1982·The Journal of Cell Biology·J J BergeronJ Paiement
Aug 1, 1981·The Journal of Cell Biology·J W Slot, H J Geuze
Jan 1, 1982·The Journal of Cell Biology·D D SabatiniM Adesnik
Jan 1, 1980·The Journal of Cell Biology·R BretzG E Palade

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Citations

Apr 1, 1991·Journal of Cellular Physiology·H BosshartE G Berger
Jan 1, 1991·Journal of Electron Microscopy Technique·M Pavelka, A Ellinger
Dec 1, 1985·The Journal of Cell Biology·H J GeuzeK von Figura
Jul 15, 1986·European Journal of Biochemistry·R P Oude ElferinkJ M Tager
Dec 1, 1987·European Journal of Biochemistry·G J StrousA L Schwartz
Nov 26, 2003·The Journal of Cell Biology·Pradipta GhoshStuart Kornfeld
Jan 1, 1986·CRC Critical Reviews in Biochemistry·G J Strous
Jan 1, 1991·Journal of Electron Microscopy Technique·H J Geuze, D J Morré
Feb 1, 1991·Journal of Electron Microscopy Technique·D M Morré
Feb 29, 1988·Biochemical and Biophysical Research Communications·G J StrousE G Berger
May 15, 1991·Biochemical and Biophysical Research Communications·L Mengle-GawD C Tiemeier
Jan 1, 1985·Journal of Cellular Biochemistry·N L ShaperJ H Shaper
Oct 5, 1987·Biochimica Et Biophysica Acta·J Roth
Oct 1, 1996·Journal of Virology·A SchlegelK Kirkegaard

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