Summary
In earlier studies from our laboratory, the intact sperm receptor was partially purified from Strongylocentrotus purpuratus crude egg membranes, but due to its insolubility, it was not possible to purify it to homogeneity. Nonetheless, this receptor preparation bound with species specificity to acrosome-reacted sperm, thereby inhibiting fertilization (17). Antibodies against the partially pure receptor inhibited fertilization in S. purpuratus (but not Arbacia punctulata) by coating the egg surface, indicating the presence of binding sites that can be species-specifically recognized by both sperm and antibody molecules.
Recently we were able to further purify and characterize the receptor from S. purpuratus eggs. Chaotropic agent solubilization of the receptor prepared from crude egg membranes yielded a very high molecular weight glycoconjugate that had many of the properties of a proteoglycan. The receptor interacted with bindin in an in vitro assay and bound with species specificity to acrosome-reacted sperm to inhibit fertilization. Unfortunately, this receptor preparation was soluble only in certain chaotropic agents (16).
Exhaustive Pronase digestion of the intact receptor yielded a soluble high-molecular-weight (>106) polysaccharide that was virtually devoid of protein. This glycosaminoglycan-like fragment was highly sulfated, and contained fucose, galactosamine, and iduronic acid (16). The fragment inhibited fertilization, but did not do so with species specificity.
Recently, soluble molecules with receptor activity were generated by treating intact dejellied eggs with trypsin. These proteolytically derived molecules contained (on a weight basis) approximately equal amounts of protein and carbohydrate. Importantly, they inhibited fertilization with species specificity. These results suggested that the binding activity was conferred by the polysaccharide component of the receptor and that the intact receptor and the tryptic fragments contained structural elements in the polypeptide chain necessary for species recognition.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Brown, A.H., 1946. Determination of pentose in the presence of large quantities of glucose. Arch. Biochem. 2: 269–278.
Conrad, G.W., Ager-Johnson, P., and Woo, M.L. 1982. Antibodies against the predominant glycosaminoglycan of the mammalian cornea, keratan sulfate. J. Biol. Chem. 257: 464–471.
Decker, G.L., and Lennarz, W.J. 1979. Sperm binding and fertilization envelope formation in a cell surface complex isolated from sea urchin eggs. J. Cell Biol. 81: 92–103.
Dische, F. 1947. A new specific color reaction of hexuronic acids. J. Biol. Chem. 167: 189–198.
Dubois, M.K., Giles, A., Hamilton, J.K., Rebers, P.A., and Smith, F. 1956. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350–356.
Foerder, C.A., and Shapiro, B.M. 1977. Release of ovoperoxidase from sea urchin eggs hardens the fertilization membrane with dityrosine crosslinks. Proc. Natl. Acad. Sci. USA 74: 4214–4218.
Fraker, P.J., and Spreck, J.C., Jr. 1978. Protein and cell membrane iodinations with a sparingly soluble chloramide, 1,3,4,6-tetrachloro-3a6a dipenyl glycoluril. Biochem. Biophys. Res. Commun. 80: 849–857.
Glabe, C.G., and Lennarz, W.J. 1979. Species-specific adhesion in sea urchins: A quantitative investigation of bindin-mediated egg agglutination. J. Cell Biol. 83: 595–604.
Glabe, C.G., and Lennarz, W.J. 1981. Isolation of a high molecular weight glycoconjugate derived from the surface of S. purpuratus eggs that is implicated in sperm adhesion. J. Supramolec. Struct. Cell Biochem. 15: 387–394.
Glabe, C.G., and Vacquier, V.D., 1977. Species-specific agglutination of eggs of bindin isolated from sea urchin sperm. Nature 267: 836–837.
Gulyas, B.J., and Schmell, E.D., 1980. Ovoperoxidase activity in ionophore treated mouse eggs. I. Electron microscopic localization. Gamete Res. 3: 267–277.
Kinsey, W.J., and Lennarz, W.J., 1981. Isolation of a glycopeptide fraction from the surface of the sea urchin egg that inhibits sperm-egg binding and fertilization. J. Cell Biol. 91: 325–331.
Kwapinski, G., 1982. In: “The Methodology of Investigative and Clinical Immunology”, Robert E. Krieger Publishing Co., Florida, pp. 173–174.
Lopo, A.C., and Vacquier, V.D., 1980. Antibody to a sperm surface glycoprotein inhibits the egg jelly induced acrosome reaction of sea urchin sperm. Dev. Biol. 79: 325–333.
Moy, G.W., and Vacquier, V.D., 1979. Immunoperoxidase localization of bindin during sea urchin fertilization. Curr. Top. Dev. Biol. 13: 31–44.
Rossignol, D.P., Earles, B.J., Decker, G.L., and Lennarz, W.J., 1984 Characterization of the sperm receptor on the surface of eggs of Strongylocentrotus purpuratus. Dev. Biol. 104: 308–3210.
Rossignol, D.P., Roschelle, A.J., and Lennarz, W.J., 1981. Sperm-egg binding: Identification of a species-specific sperm receptor from eggs of Strongylocentrotus purpuratus. J. Supramolec. Struct. Cell Biochem. 15: 347–358.
Schmell, E., Earles, B.J., Breaux, C., and Lennarz, W.J., 1977. Identification of a sperm receptor on the surface of the eggs of the sea urchin Arbacia punctulata. J. Cell Biol. 72: 35–46.
SeGall, G.K., and Lennarz, W.J., 1979. Chemical characterization of the component of the jelly coat from sea urchin eggs responsible for induction of the acrosome reaction. Dev. Biol. 71: 33–48.
SeGall, G.K., and Lennarz, W.J., 1981. Jelly coat and induction of the acrosome reaction in echinoid sperm. Dev. Biol., 86: 87–93.
Spiro, M.J., 1977. Uronic acid analysis by automated anion exchange chromatography. Anal. Biochem. 82: 348–352.
Svennerholm, L., 1956. The determination of hexosamines special references to nervous tissue. Acta. Soc. Med. Upsol. 61: 287–306.
Terho, T.T., and Hartiola, K., 1971. Method for determination of the sulfate content of glycosaminoglycans. Anal. Biochem. 41: 471–476.
Vacquier, V.D., and Moy, G.W., 1977. Isolation of bindin: The protein responsible for adhesion of sperm to sea urchin eggs. Proc. Natl. Acad. Sci. USA 74: 2456–2460.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1986 Plenum Press, New York
About this chapter
Cite this chapter
Ruiz-Bravo, N., Rossignol, D.P., Decker, G.L., Rosenberg, L.I., Lennarz, W.J. (1986). Characterization of the Strongylocentrotus Purpuratus Egg Cell Surface Receptor for Sperm. In: Hedrick, J.L. (eds) The Molecular and Cellular Biology of Fertilization. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-2255-9_17
Download citation
DOI: https://doi.org/10.1007/978-1-4613-2255-9_17
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4612-9320-0
Online ISBN: 978-1-4613-2255-9
eBook Packages: Springer Book Archive