The Journal of Membrane Biology

, Volume 19, Issue 1, pp 397–408 | Cite as

Recovery of galactosyltransferase activity from sucrose gradients during isolation of Golgi membranes

  • M. M. Mitranic
  • J. M. Sturgess
  • M. A. Moscarello


The purification of Golgi membranes has been characterized in membrane fractions isolated from rat liver by homogenization and by differential centrifugation on a discontinuous sucrose gradient. The purification of membranes, judged from the ratio of galactosyltransferase in the Golgi-rich fractions to that in the liver homogenate, correlated with the proportion of structurally intact isolated membranes. With increasing purification of membrane fractions, there was a corresponding increase in the total recovery of enzyme activity. It is postulated that this may reflect not only an increase in the purification of membranes but may involve the presence of specific galactosyltransferase inhibitors in intracellular membranes of the liver.


Sucrose Enzyme Activity Human Physiology Membrane Fraction Liver Homogenate 
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  1. Aronson, N. N., Tan, L. Y., Peters, B. P. 1973. Galactosyltransferase — The liver plasma membrane binding site for asialoglycoproteins.Biochem. Biophys. Res. Commun. 53:112PubMedGoogle Scholar
  2. Brew, K., Vanaman, T. C., Hill, R. L. 1968. The role of α-lactalbumin and the A protein in lactose synthetase: A unique mechanism for the control of a biological function.Proc. Nat. Acad. Sci. 59:491PubMedGoogle Scholar
  3. Ehrenreich, J. H., Bergeron, J. J. M., Siekevitz, P., Palade, G. E. 1973. Golgi fractions prepared from rat liver homogenates.J. Cell Biol. 59:45PubMedGoogle Scholar
  4. Fleischer, B., Fleischer, S. 1970. Preparation and characterization of Golgi membranes from rat liver.Biochim. Biophys. Acta 219:301PubMedGoogle Scholar
  5. Lawford, G. R., Schachter, H. 1966. The incorporationin vivo of14C-glucosamine into protein bound hexosamine and sialic acid of rat liver subcellular fractions.J. Biol. Chem. 241:5408PubMedGoogle Scholar
  6. Leelavathi, D. E., Estes, L. W., Feingold, D., Lombardi, B. 1970. Isolation of a Golgi rich fraction from rat liver.Biochim. Biophys. Acta 24:124Google Scholar
  7. Lowry, O. H., Rosebrough, N. J., Farr, A. G., Randall, R. J. 1951. Protein measurement with the Folin phenol reagent.J. Biol. Chem. 193:265PubMedGoogle Scholar
  8. McGuire, E. J., Jourdian, G. W., Carlson, D. M., Roseman, S. 1965. Incorporation ofd-galactose into glycoprotein.J. Biol. Chem. 240:4113Google Scholar
  9. Mookerjea, S., Michaels, M. A., Hudgin, R. L., Moscarello, M. A., Chow, A., Schachter, H. 1972. The levels of nucleotide sugar: Glycoprotein sialyl and N-acetylglucosaminyl-transferases in normal and pathological human sera.Canad. J. Biochem. 50:738Google Scholar
  10. Moore, S., Stein, W. J. 1954. A modified ninhydrin reagent for the photometric determination of amino acids and related compounds.J. Biol. Chem. 211:907PubMedGoogle Scholar
  11. Morré, D. J., Hamilton, R. L., Mollenhauer, H. H., Mahley, R. W., Cunningham, W. P., Cheetham, R. D., Lequire, V. S. 1970. Isolation of a Golgi apparatus rich fraction from rat liver.J. Cell Biol. 29:373Google Scholar
  12. Roseman, S. 1968. Biosynthesis of glycoproteins, gangliosides and related substances.In: Biochemistry of Glycoproteins and Related Substances — Cystic Fibrosis, Part II. E. Rossi and E. Stoll, editors. p. 245. P. Karger, BaselGoogle Scholar
  13. Schachter, H., Jabbal, I., Hudgin, R. L., Pinteric, L., McGuire, E. J., Roseman, S. 1970. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in Golgi-rich fractions.J. Biol. Chem. 245:1090PubMedGoogle Scholar
  14. Schachter, H., Roden, L. 1973. The biosynthesis of animal glycoproteins.In: Metabolic Conjugation and Metabolic Hydrolysis. H. Fishman, editor. Vol. III, p. 1. Academic Press, New York-LondonGoogle Scholar
  15. Sturgess, J. M., Iglesia, F. de la. 1972. Morphometry of the Golgi apparatus in developing liver.J. Cell Biol. 55:524PubMedGoogle Scholar
  16. Sturgess, J. M., Katona, E., Moscarello, M. A. 1973. The Golgi Complex. I. Isolation and ultrastructure in normal rat liver.J. Membrane Biol. 12:367Google Scholar
  17. Treloar, M. 1974. Effects of Puromycin on Galactosyltransferase of Golgi Membranes. Master of Science Thesis. University of Toronto, CanadaGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 1974

Authors and Affiliations

  • M. M. Mitranic
    • 1
  • J. M. Sturgess
    • 1
  • M. A. Moscarello
    • 1
  1. 1.Biochemistry Department and Pathology Department, Research InstituteThe Hospital for Sick ChildrenTorontoCanada

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