Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Nov 1;91(2):361–372. doi: 10.1083/jcb.91.2.361

Freeze-fracture cytochemistry: localization of wheat-germ agglutinin and concanavalin A binding sites on freeze-fractured pancreatic cells

PMCID: PMC2111992  PMID: 7031067

Abstract

The combined application of thin-section and critical-point-drying "fracture-label" is used to determine the pattern of distribution and partition of wheat-germ agglutinin and concanavalin A binding sites on the membrane faces of freeze-fractured exocrine and endocrine rat pancreatic cells. Whereas the exoplasmic face of plasma membrane is preferentially labeled by both lectins, the endoplasmic reticulum and nuclear envelope are strongly and uniformly labeled by concanavalin A but not by wheat-germ agglutinin. The results support current views in the glycosylation of membrane proteins and do not support the backflow of sialidated glycoproteins to the endoplasmic reticulum.

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

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

  1. Ackerman G. A. Distribution of wheat germ agglutinin binding sites on normal human and guinea pig bone marrow cells: an ultrastructural histochemical study. Anat Rec. 1979 Dec;195(4):641–657. doi: 10.1002/ar.1091950406. [DOI] [PubMed] [Google Scholar]
  2. Bergman A., Dallner G. Distribution of protein-bound sugar residues in microsomal subfractions and Golgi membranes. Biochim Biophys Acta. 1976 May 21;433(3):496–508. doi: 10.1016/0005-2736(76)90276-5. [DOI] [PubMed] [Google Scholar]
  3. Branton D. Fracture faces of frozen membranes. Proc Natl Acad Sci U S A. 1966 May;55(5):1048–1056. doi: 10.1073/pnas.55.5.1048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. DePierre J. W., Lazdins J., Karnovsky M. L. The determination and localization of sialic acid in guinea-pig granulocytes. Biochem J. 1980 Nov 15;192(2):543–550. doi: 10.1042/bj1920543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Elhammer A., Svensson H., Autuori F., Dallner G. Biogenesis of microsomal membrane glycoproteins in rat liver. III. Release of glycoproteins from the Golgi fraction and their transfer to microsomal membranes. J Cell Biol. 1975 Dec;67(3):715–724. doi: 10.1083/jcb.67.3.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Geoghegan W. D., Ackerman G. A. Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem. 1977 Nov;25(11):1187–1200. doi: 10.1177/25.11.21217. [DOI] [PubMed] [Google Scholar]
  8. Horisberger M., Rosset J. Colloidal gold, a useful marker for transmission and scanning electron microscopy. J Histochem Cytochem. 1977 Apr;25(4):295–305. doi: 10.1177/25.4.323352. [DOI] [PubMed] [Google Scholar]
  9. Monneron A., Segretain D. Extensive binding of concanavalin A to the nuclear membrane. FEBS Lett. 1974 Jun 1;42(2):209–213. doi: 10.1016/0014-5793(74)80787-8. [DOI] [PubMed] [Google Scholar]
  10. Parry G. Membrane assembly and turnover. Subcell Biochem. 1978;5:261–325. doi: 10.1007/978-1-4615-7942-7_6. [DOI] [PubMed] [Google Scholar]
  11. Pinto da Silva P., Branton D. Membrane splitting in freeze-ethching. Covalently bound ferritin as a membrane marker. J Cell Biol. 1970 Jun;45(3):598–605. doi: 10.1083/jcb.45.3.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pinto da Silva P., Parkison C., Dwyer N. Fracture-label:O cytochemistry of freeze-fracture faces in the erythrocyte membrane. Proc Natl Acad Sci U S A. 1981 Jan;78(1):343–347. doi: 10.1073/pnas.78.1.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Richardson C. L., Keenan T. W., Morre D. J. Ganglioside biosynthesis. Characterization of CMP-N-acetylneuraminic acid : lactosylceramide sialyltransferase in Golgi apparatus from rat liver. Biochim Biophys Acta. 1977 Jul 20;488(1):88–96. doi: 10.1016/0005-2760(77)90125-4. [DOI] [PubMed] [Google Scholar]
  14. Robbins P. W., Hubbard S. C., Turco S. J., Wirth D. F. Proposal for a common oligosaccharide intermediate in the synthesis of membrane glycoproteins. Cell. 1977 Dec;12(4):893–900. doi: 10.1016/0092-8674(77)90153-2. [DOI] [PubMed] [Google Scholar]
  15. Rodriguez Boulan E., Kreibich G., Sabatini D. D. Spatial orientation of glycoproteins in membranes of rat liver rough microsomes. I. Localization of lectin-binding sites in microsomal membranes. J Cell Biol. 1978 Sep;78(3):874–893. doi: 10.1083/jcb.78.3.874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ronzio R. A., Kronquist K. E., Lewis D. S., MacDonald R. J., Mohrlok S. H., O'Donnell J. J., Jr Glycoprotein synthesis in the adult rat pancreas. IV. Subcellular distribution of membrane glycoproteins. Biochim Biophys Acta. 1978 Mar 21;508(1):65–84. doi: 10.1016/0005-2736(78)90189-x. [DOI] [PubMed] [Google Scholar]
  17. Sesso A., Assis J. E., Kuwajima V. Y., Kachar B. Freeze-fracture and thin-section study of condensing vacuoles in rat pancreatic acinar cells. Acta Anat (Basel) 1980;108(4):521–539. doi: 10.1159/000145351. [DOI] [PubMed] [Google Scholar]
  18. Snider M. D., Sultzman L. A., Robbins P. W. Transmembrane location of oligosaccharide-lipid synthesis in microsomal vesicles. Cell. 1980 Sep;21(2):385–392. doi: 10.1016/0092-8674(80)90475-4. [DOI] [PubMed] [Google Scholar]
  19. Svensson H., Elhammer A., Autuori F., Dallner G. Biogenesis of microsomal membrane glycoproteins in rat liver. IV. Characteristics of a cytoplasmic lipoprotein having properties of a membrane precursor. Biochim Biophys Acta. 1976 Dec 2;455(2):383–398. doi: 10.1016/0005-2736(76)90313-8. [DOI] [PubMed] [Google Scholar]
  20. Virtanen I., Ekblom P., Laurila P. Subcellular compartmentalization of saccharide moieties in cultured normal and malignant cells. J Cell Biol. 1980 May;85(2):429–434. doi: 10.1083/jcb.85.2.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Virtanen I. Identification of concanavalin A binding glycoproteins of rat liver cell nuclear membranes. Biochem Biophys Res Commun. 1977 Oct 24;78(4):1411–1417. doi: 10.1016/0006-291x(77)91449-8. [DOI] [PubMed] [Google Scholar]
  22. Virtanen I., Wartiovaara J. Lectin receptor sites on rat liver cell nuclear membranes. J Cell Sci. 1976 Nov;22(2):335–344. doi: 10.1242/jcs.22.2.335. [DOI] [PubMed] [Google Scholar]
  23. Winqvist L., Eriksson L. C., Dallner G. Interaction of lectins with proteins of the endoplasmic reticulum and Golgi system of rat liver. J Cell Sci. 1979 Oct;39:101–116. doi: 10.1242/jcs.39.1.101. [DOI] [PubMed] [Google Scholar]
  24. da Silva P. P., Kachar B., Torrisi M. R., Brown C., Parkison C. Freeze-fracture cytochemistry: replicas of critical point-dried cells and tissues after fracture-label. Science. 1981 Jul 10;213(4504):230–233. doi: 10.1126/science.7244630. [DOI] [PubMed] [Google Scholar]
  25. da Silva P. P., Parkison C., Dwyer N. Freeze-fracture cytochemistry: thin sections of cells and tissues after labeling of fractures faces. J Histochem Cytochem. 1981 Aug;29(8):917–928. doi: 10.1177/29.8.7276536. [DOI] [PubMed] [Google Scholar]

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

RESOURCES