Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1972 Oct;69(10):2945–2949. doi: 10.1073/pnas.69.10.2945

Distribution of Saccharide Residues on Membrane Fragments from a Myeloma-Cell Homogenate: Its Implications for Membrane Biogenesis

Hiroshi Hirano *,†, Brenda Parkhouse ‡,§, Garth L Nicolson ‡, Edwin S Lennox ‡, S J Singer *,
PMCID: PMC389680  PMID: 4117011

Abstract

Ferritin conjugates of two plant agglutinins, concanavalin A and ricin, have been used as specific electron microscopic stains for covalently-bound saccharide residues on membrane fragments from a myeloma-cell homogenate. The results indicate that different saccharide residues are uniformly localized to a single surface of each membrane fragment. In particular, the ferritin-concanavalin A conjugate binds exclusively to the cisternal side of membrane fragments of the rough endoplasmic reticulum. If it is postulated that the biogenesis of eukaryotic plasma membranes involves an assembly-line process from precursor intracellular membranes, these observed asymmetric distributions of saccharides on cell membranes can be explained.

Keywords: ferritin conjugates, plant agglutinins, smooth- and rough-membrane elements

Full text

PDF
2945

Images in this article

2946Image
on p.2946
2947Image
on p.2947

Selected References

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

  1. Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
  2. Bennett G., Leblond C. P. Formation of cell coat material for the whole surface of columnar cells in the rat small intestine, as visualized by radioautography with L-fucose-3H. J Cell Biol. 1970 Aug;46(2):409–416. doi: 10.1083/jcb.46.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Choi Y. S., Knopf P. M., Lennox E. S. Intracellular transport and secretion of an immunoglobulin light chain. Biochemistry. 1971 Feb 16;10(4):668–679. doi: 10.1021/bi00780a019. [DOI] [PubMed] [Google Scholar]
  4. Choi Y. S., Knopf P. M., Lennox E. S. Subcellular fractionation of mouse myeloma cells. Biochemistry. 1971 Feb 16;10(4):659–667. doi: 10.1021/bi00780a018. [DOI] [PubMed] [Google Scholar]
  5. Drysdale R. G., Herrick P. R., Franks D. The specificity of the haemagglutinin of the Castor bean, Ricinus communis. Vox Sang. 1968;15(3):194–202. doi: 10.1111/j.1423-0410.1968.tb01749.x. [DOI] [PubMed] [Google Scholar]
  6. Frye L. D., Edidin M. The rapid intermixing of cell surface antigens after formation of mouse-human heterokaryons. J Cell Sci. 1970 Sep;7(2):319–335. doi: 10.1242/jcs.7.2.319. [DOI] [PubMed] [Google Scholar]
  7. Horibata K., Harris A. W. Mouse myelomas and lymphomas in culture. Exp Cell Res. 1970 Apr;60(1):61–77. doi: 10.1016/0014-4827(70)90489-1. [DOI] [PubMed] [Google Scholar]
  8. Kornberg R. D., McConnell H. M. Inside-outside transitions of phospholipids in vesicle membranes. Biochemistry. 1971 Mar 30;10(7):1111–1120. doi: 10.1021/bi00783a003. [DOI] [PubMed] [Google Scholar]
  9. Marchesi V. T., Palade G. E. The localization of Mg-Na-K-activated adenosine triphosphatase on red cell ghost membranes. J Cell Biol. 1967 Nov;35(2):385–404. doi: 10.1083/jcb.35.2.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Molnar J., Sy D. Attachment of glucosamine to protein at the ribosomal site of rat liver. Biochemistry. 1967 Jul;6(7):1941–1947. doi: 10.1021/bi00859a009. [DOI] [PubMed] [Google Scholar]
  11. Nicolson G. L., Blaustein J. The interaction of Ricinus communis agglutinin with normal and tumor cell surfaces. Biochim Biophys Acta. 1972 May 9;266(2):543–547. doi: 10.1016/0005-2736(72)90109-5. [DOI] [PubMed] [Google Scholar]
  12. Nicolson G. L., Marchesi V. T., Singer S. J. The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies. J Cell Biol. 1971 Oct;51(1):265–272. doi: 10.1083/jcb.51.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nicolson G. L., Singer S. J. Ferritin-conjugated plant agglutinins as specific saccharide stains for electron microscopy: application to saccharides bound to cell membranes. Proc Natl Acad Sci U S A. 1971 May;68(5):942–945. doi: 10.1073/pnas.68.5.942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Poretz R. D., Goldstein I. J. An examination of the topography of the saccharide binding sites of concanavalin A and of the forces involved in complexation. Biochemistry. 1970 Jul 7;9(14):2890–2896. doi: 10.1021/bi00816a021. [DOI] [PubMed] [Google Scholar]
  15. Rambourg A., Hernandez W., Leblond C. P. Detection of complex carbohydrates in the Golgi apparatus of rat cells. J Cell Biol. 1969 Feb;40(2):395–414. doi: 10.1083/jcb.40.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rambourg A. Morphological and histochemical aspects of glycoproteins at the surface of animal cells. Int Rev Cytol. 1971;31:57–114. doi: 10.1016/s0074-7696(08)60057-1. [DOI] [PubMed] [Google Scholar]
  17. Ray T. K., Lieberman I., Lansing A. I. Synthesis of the plasma membrane of the liver cell. Biochem Biophys Res Commun. 1968 Apr 5;31(1):54–58. doi: 10.1016/0006-291x(68)90030-2. [DOI] [PubMed] [Google Scholar]
  18. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  19. Spiro R. G. Glycoproteins. Annu Rev Biochem. 1970;39:599–638. doi: 10.1146/annurev.bi.39.070170.003123. [DOI] [PubMed] [Google Scholar]
  20. Stockem W. Pinocytose und Bewegung von Amöben. 3. Die Funktsion des Golgiapparates von Amoeba proteus und Chaos chaos. Histochemie. 1969;18(3):217–240. doi: 10.1007/BF00306169. [DOI] [PubMed] [Google Scholar]
  21. Whur P., Herscovics A., Leblond C. P. Radioautographic visualization of the incorporation of galactose-3H and mannose-3H by rat thyroids in vitro in relation to the stages of thyroglobulin synthesis. J Cell Biol. 1969 Nov;43(2):289–311. doi: 10.1083/jcb.43.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wise G. E., Flickinger C. J. Cytochemical staining of the Golgi apparatus in amoeba proteus. J Cell Biol. 1970 Sep;46(3):620–626. doi: 10.1083/jcb.46.3.620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wise G. E., Flickinger C. J. Relation of the Golgi apparatus to the cell coat in amoebae. Exp Cell Res. 1970 Jul;61(1):13–23. doi: 10.1016/0014-4827(70)90252-1. [DOI] [PubMed] [Google Scholar]
  24. Zagury D., Uhr J. W., Jamieson J. D., Palade G. E. Immunoglobulin synthesis and secretion. II. Radioautographic studies of sites of addition of carbohydrate moieties and intracellular transport. J Cell Biol. 1970 Jul;46(1):52–63. doi: 10.1083/jcb.46.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES