Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Nov 1;99(5):1598–1604. doi: 10.1083/jcb.99.5.1598

Wheat germ agglutinin blocks the acrosome reaction in Strongylocentrotus purpuratus sperm by binding a 210,000-mol-wt membrane protein

PMCID: PMC2113364  PMID: 6092388

Abstract

Wheat germ agglutinin (WGA) binds to the entire surface of Strongylocentrotus purpuratus sperm, and inhibits the egg jelly-induced acrosome reaction. The binding was found to be species dependent and was completely inhibited by 5 mM N-acetyl-D-glucosamine. Blockage of the acrosome reaction by WGA was bypassed by a combination of the ionophores A23187 and monensin, although neither ionophore was effective individually. These experiments suggest that WGA blocks both Ca2+ uptake and Na+/H+ exchange in these sperm, which was confirmed by direct measurements of 45Ca2+ uptake and H+ efflux. The target of WGA in S. purpuratus sperm appears to be a membrane glycoprotein of Mr = 210,000. Treatment of this protein with neuraminidase or endo-beta-N- acetylglucosaminidase F abolished WGA binding.

Full Text

The Full Text of this article is available as a PDF (711.2 KB).

Selected References

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

  1. Bhavanandan V. P., Katlic A. W. The interaction of wheat germ agglutinin with sialoglycoproteins. The role of sialic acid. J Biol Chem. 1979 May 25;254(10):4000–4008. [PubMed] [Google Scholar]
  2. Cross N. L. Isolation and electrophoretic characterization of the plasma membrane of sea-urchin sperm. J Cell Sci. 1983 Jan;59:13–25. doi: 10.1242/jcs.59.1.13. [DOI] [PubMed] [Google Scholar]
  3. Eckberg W. R., Metz C. B. Isolation of an arbacia sperm fertilization antigen. J Exp Zool. 1982 May 20;221(1):101–105. doi: 10.1002/jez.1402210113. [DOI] [PubMed] [Google Scholar]
  4. Elder J. H., Alexander S. endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4540–4544. doi: 10.1073/pnas.79.15.4540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feramisco J. R. Microinjection of fluorescently labeled alpha-actinin into living fibroblasts. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3967–3971. doi: 10.1073/pnas.76.8.3967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Lee H. C., Johnson C., Epel D. Changes in internal pH associated with initiation of motility and acrosome reaction of sea urchin sperm. Dev Biol. 1983 Jan;95(1):31–45. doi: 10.1016/0012-1606(83)90004-0. [DOI] [PubMed] [Google Scholar]
  8. Lopo A. C., Vacquier V. D. Antibody to a sperm surface glycoprotein inhibits the egg jelly-induced acrosome reaction of sea urchin sperm. Dev Biol. 1980 Oct;79(2):325–333. doi: 10.1016/0012-1606(80)90119-0. [DOI] [PubMed] [Google Scholar]
  9. Maylié-Pfenninger M. F., Jamieson J. D. Distribution of cell surface saccharides on pancreatic cells. I. General method for preparation and purification of lectins and lectin-ferritin conjugates. J Cell Biol. 1979 Jan;80(1):69–76. doi: 10.1083/jcb.80.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Monsigny M., Sene C., Obrenovitch A., Roche A. C., Delmotte F., Boschetti E. Properties of succinylated wheat-germ agglutinin. Eur J Biochem. 1979 Jul;98(1):39–45. doi: 10.1111/j.1432-1033.1979.tb13157.x. [DOI] [PubMed] [Google Scholar]
  11. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  12. Muresan V., Iwanij V., Smith Z. D., Jamieson J. D. Purification and use of limulin: a sialic acid-specific lectin. J Histochem Cytochem. 1982 Sep;30(9):938–946. doi: 10.1177/30.9.6897073. [DOI] [PubMed] [Google Scholar]
  13. Peters B. P., Ebisu S., Goldstein I. J., Flashner M. Interaction of wheat germ agglutinin with sialic acid. Biochemistry. 1979 Nov 27;18(24):5505–5511. doi: 10.1021/bi00591a038. [DOI] [PubMed] [Google Scholar]
  14. Saling P. M., Eckberg W. R., Metz C. B. Mechanism of univalent antisperm antibody inhibition of fertilization in the sea urchin, Arbacia punctulata. J Exp Zool. 1982 May 20;221(1):93–99. doi: 10.1002/jez.1402210112. [DOI] [PubMed] [Google Scholar]
  15. Schackmann R. W., Christen R., Shapiro B. M. Membrane potential depolarization and increased intracellular pH accompany the acrosome reaction of sea urchin sperm. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6066–6070. doi: 10.1073/pnas.78.10.6066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schackmann R. W., Shapiro B. M. A partial sequence of ionic changes associated with the acrosome reaction of Strongylocentrotus purpuratus. Dev Biol. 1981 Jan 15;81(1):145–154. doi: 10.1016/0012-1606(81)90357-2. [DOI] [PubMed] [Google Scholar]
  17. SeGall G. K., Lennarz W. J. Chemical characterization of the component of the jelly coat from sea urchin eggs responsible for induction of the acrosome reaction. Dev Biol. 1979 Jul;71(1):33–48. doi: 10.1016/0012-1606(79)90080-0. [DOI] [PubMed] [Google Scholar]
  18. SeGall G. K., Lennarz W. J. Jelly coat and induction of the acrosome reaction in echinoid sperm. Dev Biol. 1981 Aug;86(1):87–93. doi: 10.1016/0012-1606(81)90318-3. [DOI] [PubMed] [Google Scholar]
  19. Tilney L. G., Hatano S., Ishikawa H., Mooseker M. S. The polymerization of actin: its role in the generation of the acrosomal process of certain echinoderm sperm. J Cell Biol. 1973 Oct;59(1):109–126. doi: 10.1083/jcb.59.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vacquier V. D., Payne J. E. Methods for quantitating sea urchin sperm-egg binding. Exp Cell Res. 1973 Nov;82(1):227–235. doi: 10.1016/0014-4827(73)90265-6. [DOI] [PubMed] [Google Scholar]
  21. Vretblad P. Purification of lectins by biospecific affinity chromatography. Biochim Biophys Acta. 1976 May 20;434(1):169–176. doi: 10.1016/0005-2795(76)90047-7. [DOI] [PubMed] [Google Scholar]
  22. Wright C. S. Location of the N-acetyl-D-neuraminic acid binding site in wheat germ agglutinin. A crystallographic study at 2.8 A resolution. J Mol Biol. 1980 May 5;139(1):53–60. doi: 10.1016/0022-2836(80)90115-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES