Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Jul 1;97(1):270–275. doi: 10.1083/jcb.97.1.270

Compartmentation of asparagine-linked oligosaccharide processing in the Golgi apparatus

PMCID: PMC2112488  PMID: 6223041

Abstract

Golgi-associated processing of complex-type oligosaccharides linked to asparagine involves the sequential action of at least six enzymes. By equilibrium sucrose density gradient centrifugation of membranes from Chinese hamster ovary cells, we have partially resolved the set of four initial enzymes in the pathway (Mannosidase I, N-acetylglucosamine (GlcNAc) Transferase I, Mannosidase II, and GlcNAc Transferase II) from two later-acting activities (galactosyltransferase and sialyltransferase). In view of the recent demonstration that galactosyltransferase is restricted to the trans face of the Golgi complex in HeLa cells (Roth, J., and E.G. Berger, 1982, J. Cell Biol., 93:223-229), our results suggest that removal of mannose and attachment of peripheral N-acetylglucosamine may occur in some or all of the remaining cisternae on the cis side of the Golgi stack.

Full Text

The Full Text of this article is available as a PDF (606.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bergeron J. J., Ehrenreich J. H., Siekevitz P., Palade G. E. Golgi fractions prepared from rat liver homogenates. II. Biochemical characterization. J Cell Biol. 1973 Oct;59(1):73–88. doi: 10.1083/jcb.59.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borgese N., Meldolesi J. Localization and biosynthesis of NADH-cytochrome b5 reductase, an integral membrane protein, in rat liver cells. I. Distribution of the enzyme activity in microsomes, mitochondria, and golgi complex. J Cell Biol. 1980 Jun;85(3):501–515. doi: 10.1083/jcb.85.3.501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bretz R., Bretz H., Palade G. E. Distribution of terminal glycosyltransferases in hepatic Golgi fractions. J Cell Biol. 1980 Jan;84(1):87–101. doi: 10.1083/jcb.84.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Briles E. B., Li E., Kornfeld S. Isolation of wheat germ agglutinin-resistant clones of Chinese hamster ovary cells deficient in membrane sialic acid and galactose. J Biol Chem. 1977 Feb 10;252(3):1107–1116. [PubMed] [Google Scholar]
  5. Cheetham R. D., Morré D. J., Pannek C., Friend D. S. Isolation of a Golgi apparatus-rich fraction from rat liver. IV. Thiamine pyrophosphatase. J Cell Biol. 1971 Jun;49(3):899–905. doi: 10.1083/jcb.49.3.899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ehrenreich J. H., Bergeron J. J., Siekevitz P., Palade G. E. Golgi fractions prepared from rat liver homogenates. I. Isolation procedure and morphological characterization. J Cell Biol. 1973 Oct;59(1):45–72. doi: 10.1083/jcb.59.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FRIEND D. S., MURRAY M. J. OSMIUM IMPREGNATION OF THE GOLGI APPARATUS. Am J Anat. 1965 Jul;117:135–149. doi: 10.1002/aja.1001170109. [DOI] [PubMed] [Google Scholar]
  8. Farquhar M. G., Palade G. E. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage. J Cell Biol. 1981 Dec;91(3 Pt 2):77s–103s. doi: 10.1083/jcb.91.3.77s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fleischer B., Fleischer S., Ozawa H. Isolation and characterization of Golgi membranes from bovine liver. J Cell Biol. 1969 Oct;43(1):59–79. doi: 10.1083/jcb.43.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldberg D. E., Kornfeld S. Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation. J Biol Chem. 1983 Mar 10;258(5):3159–3165. [PubMed] [Google Scholar]
  11. Goldberg D. E., Kornfeld S. The phosphorylation of beta-glucuronidase oligosaccharides in mouse P388D1 cells. J Biol Chem. 1981 Dec 25;256(24):13060–13067. [PubMed] [Google Scholar]
  12. Gottlieb C., Baenziger J., Kornfeld S. Deficient uridine diphosphate-N-acetylglucosamine:glycoprotein N-acetylglucosaminyltransferase activity in a clone of Chinese hamster ovary cells with altered surface glycoproteins. J Biol Chem. 1975 May 10;250(9):3303–3309. [PubMed] [Google Scholar]
  13. Griffiths G., Brands R., Burke B., Louvard D., Warren G. Viral membrane proteins acquire galactose in trans Golgi cisternae during intracellular transport. J Cell Biol. 1982 Dec;95(3):781–792. doi: 10.1083/jcb.95.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Grinna L. S., Robbins P. W. Glycoprotein biosynthesis. Rat liver microsomal glucosidases which process oligosaccharides. J Biol Chem. 1979 Sep 25;254(18):8814–8818. [PubMed] [Google Scholar]
  15. Grinna L. S., Robbins P. W. Substrate specificities of rat liver microsomal glucosidases which process glycoproteins. J Biol Chem. 1980 Mar 25;255(6):2255–2258. [PubMed] [Google Scholar]
  16. Hasilik A., Von Figura K. Oligosaccharides in lysosomal enzymes. Distribution of high-mannose and complex oligosaccharides in cathepsin D and beta-hexosaminidase. Eur J Biochem. 1981 Dec;121(1):125–129. doi: 10.1111/j.1432-1033.1981.tb06440.x. [DOI] [PubMed] [Google Scholar]
  17. Hino Y., Asano A., Sato R. Biochemical studies on rat liver Golgi apparatus. III. Subfractionation of fragmented Golgi apparatus by counter-current distribution. J Biochem. 1978 Apr;83(4):935–942. doi: 10.1093/oxfordjournals.jbchem.a132020. [DOI] [PubMed] [Google Scholar]
  18. Hubbard S. C., Ivatt R. J. Synthesis and processing of asparagine-linked oligosaccharides. Annu Rev Biochem. 1981;50:555–583. doi: 10.1146/annurev.bi.50.070181.003011. [DOI] [PubMed] [Google Scholar]
  19. Hubbard S. C., Robbins P. W. Synthesis and processing of protein-linked oligosaccharides in vivo. J Biol Chem. 1979 Jun 10;254(11):4568–4576. [PubMed] [Google Scholar]
  20. Ito A., Palade G. E. Presence of NADPH-cytochrome P-450 reductase in rat liver Golgi membranes. Evidence obtained by immunoadsorption method. J Cell Biol. 1978 Nov;79(2 Pt 1):590–597. doi: 10.1083/jcb.79.2.590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kornfeld S., Li E., Tabas I. The synthesis of complex-type oligosaccharides. II. Characterization of the processing intermediates in the synthesis of the complex oligosaccharide units of the vesicular stomatitis virus G protein. J Biol Chem. 1978 Nov 10;253(21):7771–7778. [PubMed] [Google Scholar]
  22. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  23. Li E., Tabas I., Kornfeld S. The synthesis of complex-type oligosaccharides. I. Structure of the lipid-linked oligosaccharide precursor of the complex-type oligosaccharides of the vesicular stomatitis virus G protein. J Biol Chem. 1978 Nov 10;253(21):7762–7770. [PubMed] [Google Scholar]
  24. Li S. C., Mazzotta M. Y., Chien S. F., Li Y. T. Isolation and characterization of jack bean beta-galactosidase. J Biol Chem. 1975 Sep 10;250(17):6786–6791. [PubMed] [Google Scholar]
  25. Narasimhan S., Stanley P., Schachter H. Control of glycoprotein synthesis. Lectin-resistant mutant containing only one of two distinct N-acetylglucosaminyltransferase activities present in wild type Chinese hamster ovary cells. J Biol Chem. 1977 Jun 10;252(11):3926–3933. [PubMed] [Google Scholar]
  26. Oppenheimer C. L., Hill R. L. Purification and characterization of a rabbit liver alpha 1 goes to 3 mannoside beta 1 goes to 2 N-acetylglucosaminyltransferase. J Biol Chem. 1981 Jan 25;256(2):799–804. [PubMed] [Google Scholar]
  27. Orci L., Montesano R., Meda P., Malaisse-Lagae F., Brown D., Perrelet A., Vassalli P. Heterogeneous distribution of filipin--cholesterol complexes across the cisternae of the Golgi apparatus. Proc Natl Acad Sci U S A. 1981 Jan;78(1):293–297. doi: 10.1073/pnas.78.1.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pohlmann R., Waheed A., Hasilik A., von Figura K. Synthesis of phosphorylated recognition marker in lysosomal enzymes is located in the cis part of Golgi apparatus. J Biol Chem. 1982 May 25;257(10):5323–5325. [PubMed] [Google Scholar]
  29. Prieels J. P., Pizzo S. V., Glasgow L. R., Paulson J. C., Hill R. L. Hepatic receptor that specifically binds oligosaccharides containing fucosyl alpha1 leads to 3 N-acetylglucosamine linkages. Proc Natl Acad Sci U S A. 1978 May;75(5):2215–2219. doi: 10.1073/pnas.75.5.2215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roth J., Berger E. G. Immunocytochemical localization of galactosyltransferase in HeLa cells: codistribution with thiamine pyrophosphatase in trans-Golgi cisternae. J Cell Biol. 1982 Apr;93(1):223–229. doi: 10.1083/jcb.93.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rothman J. E. The golgi apparatus: two organelles in tandem. Science. 1981 Sep 11;213(4513):1212–1219. doi: 10.1126/science.7268428. [DOI] [PubMed] [Google Scholar]
  32. Schachter H., Jabbal I., Hudgin R. L., Pinteric L., McGuire E. J., Roseman S. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in a Golgi-rich fraction. J Biol Chem. 1970 Mar 10;245(5):1090–1100. [PubMed] [Google Scholar]
  33. Schmidt M. F., Schlesinger M. J. Fatty acid binding to vesicular stomatitis virus glycoprotein: a new type of post-translational modification of the viral glycoprotein. Cell. 1979 Aug;17(4):813–819. doi: 10.1016/0092-8674(79)90321-0. [DOI] [PubMed] [Google Scholar]
  34. Sefton B. M. Immediate glycosylation of Sindbis virus membrane proteins. Cell. 1977 Apr;10(4):659–668. doi: 10.1016/0092-8674(77)90099-x. [DOI] [PubMed] [Google Scholar]
  35. Spiro R. G. Studies on fetuin, a glycoprotein of fetal serum. I. Isolation, chemical composition, and physiochemical properties. J Biol Chem. 1960 Oct;235(10):2860–2869. [PubMed] [Google Scholar]
  36. Tabas I., Kornfeld S. Purification and characterization of a rat liver Golgi alpha-mannosidase capable of processing asparagine-linked oligosaccharides. J Biol Chem. 1979 Nov 25;254(22):11655–11663. [PubMed] [Google Scholar]
  37. Tabas I., Kornfeld S. The synthesis of complex-type oligosaccharides. III. Identification of an alpha-D-mannosidase activity involved in a late stage of processing of complex-type oligosaccharides. J Biol Chem. 1978 Nov 10;253(21):7779–7786. [PubMed] [Google Scholar]
  38. Tabas I., Schlesinger S., Kornfeld S. Processing of high mannose oligosaccharides to form complex type oligosaccharides on the newly synthesized polypeptides of the vesicular stomatitis virus G protein and the IgG heavy chain. J Biol Chem. 1978 Feb 10;253(3):716–722. [PubMed] [Google Scholar]
  39. Tulsiani D. R., Hubbard S. C., Robbins P. W., Touster O. alpha-D-Mannosidases of rat liver Golgi membranes. Mannosidase II is the GlcNAcMAN5-cleaving enzyme in glycoprotein biosynthesis and mannosidases Ia and IB are the enzymes converting Man9 precursors to Man5 intermediates. J Biol Chem. 1982 Apr 10;257(7):3660–3668. [PubMed] [Google Scholar]
  40. Tulsiani D. R., Opheim D. J., Touster O. Purification and characterization of alpha-D-mannosidase from rat liver golgi membranes. J Biol Chem. 1977 May 25;252(10):3227–3233. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES