Carbohydrate-Binding Proteins Involved in Gamete Interaction in the Pig

  • Juan José Calvete
  • Libia Sanz
  • Edda Töpfer-Petersen
Part of the Schering Foundation Workshop book series (SCHERING FOUND, volume 4)


An essential event in the complex, mulitstep process of mammalian species-specific fertilization is gamete recognition through specific complementary molecules located on the external surfaces of the spermatozoon and the oocyte (Lillie 1913; Yanagimachi 1988). Increasing evidence points out that sperm-egg recognitions and binding actually involve carbohydrate-protein interaction (Ahuja 1982; Huang et al. 1982; Macek and Shur 1988; Jones and Williams 1990; Miller and Ax 1990). Remarkably, this interaction has been conserved in the process of fertilization throughout the whole evolutionary scale (Dale 1991). Thus, inhibition experiments with a variety of polysaccharides show that mannose and fucose contain egg surface components of the marine brown alga Fucus, which interact with complementary carbohydrate-binding structures on the sperm cell surface (Bolwell et al. 1979). Similarly, Hoshi (1984) has presented evidence suggesting that sperm-egg binding in ascidians may be mediated by the enzyme a-fucosidase located on the sperm head. Moreover, the freshwater bivalve Unio spermatozoa bind to its homologous egg at a part of the oocyte surface that binds lectins, suggesting that the sperm receptors are also fucosylcontaining glycoproteins (Focarelli et al. 1988). In the sea urchin, the sperm specific protein, bindin, a 24 kDA major protein (Gao et al. 1986) associated with the inner acrosomal membrane and exposed during the acrosome reaction, mediates the adhesion to the egg’s vitelline layer by a lectin-like activity (Vacquier and Moy 1977; Glabe et al. 1982). Bindin has a strong affinity for sulfated, fucose-containing carbohydrate structures on proteoglycans of the egg surface (De Angelis and Gabel 1987). Sulfate esters of the vitelline coat and arginine residues of the sperm protein appear to be critical for the binding (De Angelis and Gable 1988; 1990).


Seminal Plasma Zona Pellucida Acrosome Reaction Sperm Capacitation Sperm Plasma Membrane 
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  1. Aarons D, Boettger-Tong H, Holt G, Poirier GR ( 1991 Acrsome reaction induced by immunoaggregation of a proteinase inhibitor bound to the murin sperm head. Molec Reprod Develop 30: 258–264CrossRefGoogle Scholar
  2. Ahuja KK (1982) Fertilization studies in the hamster. The role of cell-surface carbohydrates. Exp Cell Res 140: 353–363PubMedCrossRefGoogle Scholar
  3. Baba T, Kashiwabara S, Watanabe K, Itoh H, Michikawa Y, Kimura K, Takada M, Fukamizu A, Arai Y (1989) Activation and maturation mechanisms of boar acrosin zymogen based on the deduced primary structure. J Biol Chem 264: 11920–11927PubMedGoogle Scholar
  4. Barkalow FJB, Schwarzbauer JE (1991) Localization of the major heparin-binding site in fibronectin. J Biol Chem 266: 7812–7818PubMedGoogle Scholar
  5. Benau DA, Storey BT (1987) Characterization of the mouse sperm plasma membrane zona-binding site sensitive to trypsin inhibitors. Biol Reprod 236: 282–292CrossRefGoogle Scholar
  6. Bleil JD, Wassarman PM (1986) Autoradiographic visualization of the mouse egg’s sperm receptor bound to sperm. J Cell Biol 102: 1363–1371PubMedCrossRefGoogle Scholar
  7. Bleil JD, Wassarman PM (1988) Galactose at the nonreducing terminus of O-linked oligosaccharides of mouse egg zona pellucida glycoprotein ZP3 is essential for the glycoprotein’s sperm receptor activity. Proc Natl Acad Sci USA 85: 6778–6782PubMedCrossRefGoogle Scholar
  8. Bleil JD, Wassarman PM (1989) Identification of a mouse sperm protein that recognizes ZP3. J Cell Biol 109: 125aGoogle Scholar
  9. Bleil JD (1991) Sperm receptors of mammalian eggs. In: Wassarman PM (ed) Elements of mammalian fertilization, vol. 1 Basic concepts. CRC Press Boca Raton, Florida, pp 133–151Google Scholar
  10. Bolwell G, Callow J, Callow M, Evans L (1979) Fertilization in brown algae. 11. Evidence for lectin sensitive complementary receptors involved in gamete recognition in Fucus serratus. J Cell Sci 36: 19–30PubMedGoogle Scholar
  11. Brown CR, Jones R (1987) Binding of zona pellucida proteins to a boar sperm prolypeptide of Mr 53000 and identification of zona moieties involved. Development 99: 333–339PubMedGoogle Scholar
  12. Cechova D, Töpfer-Petersen E, Henschen A (1988) Boar proacrosin is a single-chain molecule which has the N-terminus of the acrosin A-chain (light chain). FEBS Lett 241: 136–140PubMedCrossRefGoogle Scholar
  13. Chamberlain ME, Dean J (1989) Genomic organization of a sex specific gene: the primary sperm receptor of the mouse zona pellucida. Dev Biol 131: 207–214CrossRefGoogle Scholar
  14. Chamberlain ME, Dean J (1990) Human homology of the mouse sperm receptor. Proc Natl Acad Scie USA 87: 6014–6018CrossRefGoogle Scholar
  15. Chandonnet L, Roberts KD, Chapdelaine A, Manjunath P (1990) Identification of heparin-binding proteins in bovine seminal plasma. Molec Reprod Develop 26: 313–318CrossRefGoogle Scholar
  16. Dale B (1991) Mechanism of fertilization: plants to humans. In: Neuhoff V, Friend J (eds) Cell to cell signals in plants and animals. NATO ASI Series, vol. H51, Springer Verlag Berlin, pp 83–90CrossRefGoogle Scholar
  17. DeAngelis PL, Glabe CG (1987) Polysaccharide structural featrues that are critical for the binding of sulfated fucans to bindin, the adhesive protein from sea urchin sperm. J Biol Chem 262: 13946–13952PubMedGoogle Scholar
  18. DeAngelis PL, Glabe CG (1988) Role of basic amino acids in the interaction of bindin with sulfated fucan. Biochem 27: 8189–8194CrossRefGoogle Scholar
  19. DeAngelis PL, Glabe CG (1990) Specific recognition of sulfate esters by bindin, a sperm adhesion protein from sea urchins. Biochem Biophys Acta 1037: 100–105PubMedCrossRefGoogle Scholar
  20. Florman HM, First NL (1988a) The regulation of acrsomal exocytosis: Sperm capacitation is required for the induction of the acrosome reaction by the bovine zona pellucida in vitro. Dev Biol 128: 453–463PubMedCrossRefGoogle Scholar
  21. Florman HM, First NL (1988b) Regulation of acrosomal exocytosis: The zona pellucida-induced acrosome reaction of bovine spermatozoa is controlled by extrinsic positive regulatory elements. Dev Biol 128: 464–473PubMedCrossRefGoogle Scholar
  22. Florman HM, Babcock DF (1991) Progress toward understanding the molecular basis of capacitation. In: Wassarman PM (ed) Elements of mammalian fertilization, vol. 1, Basic concepts. CRC Press, Boca Raton, pp 105–132Google Scholar
  23. Focarelli R, Renieri T, Rosati F (1988) Polarized site of sperm entrance in the egg of a freshwater bivalve, Unio elongatulus. Dev Biol 127: 443–451CrossRefGoogle Scholar
  24. Fock-Nüzel R, Lottspeich F, Henschen A, Muller-Esterl W (1984) Boar acrosin is a two-chain molecule. Isolation and primary structure of the light chain; homology with the pro-part of other serine proteinases. Eur J Biochem 141: 441–446PubMedCrossRefGoogle Scholar
  25. Friess AE, Töpfer-Petersen E, Schill W-B (1987a) Electron microscopic localization of a fucose-binding protein in acrosome-reacted spermatozoa by the fucosyl-peroxidase-gold-method. Histochem 86: 297–303CrossRefGoogle Scholar
  26. Friess AE, Töpfer-Petersen E, Schill W-B (1987b) Fracture labeling of boar spermatozoa for the fucose-binding protein ( FBP ). Histochem 87: 181–183CrossRefGoogle Scholar
  27. Gao B, Klein Le, Britten RI, Davidson EH (1986) Sequence of mRNA coding for bindin, a species specific sea urchin sperm protein required for fertilization. Proc Natl Acad Sci USA 83: 8634–8638PubMedCrossRefGoogle Scholar
  28. Glabe CG, Grabel LB, Vacquier VD, Rosen SD (1982) Carbohydrate specificity of sea urchin sperm binding: Cell surface lectin mediating sperm-egg adhesion. J Cell Biol 94: 123–128PubMedCrossRefGoogle Scholar
  29. Hanging M, Tai-Ying Y, Zhao-Wen S (1991) Isolation, characterization, and localization of the zona pellucida binding proteins of boar sperm. Molec Reprod Develop 28: 124–130CrossRefGoogle Scholar
  30. Hoshi M (1984) Roles of sperm glycosidases and proteases in the ascidian fertilization. In: Engels et al. (eds) Advances in invertebrate reproduction, vol. 3, Elsevier, Amsterdam pp 27–40Google Scholar
  31. Huang TTFjr, Ohzu E, Yanagimachi R (1982) Evidence suggesting that L-fucose is part of a recognition signal for sperm-zona pellucida attachment in mammals. Gamete Res 5: 355–361CrossRefGoogle Scholar
  32. Jaikaria NS, Rosenfeld L, Khan MY, Danishewsky I, Newman SA (1991) Interaction fo fibronectin with heparin in model extracellular matrices. Role of arginine residues and sulfate groups. Biochem 30: 1538–1544CrossRefGoogle Scholar
  33. Jonökovâ V, Sanz L, Calvete JJ, Henschen A, Cechovâ D, Töpfer-Petersen E (1991) Isolation and biochemical characterization of a zona pellucida-binding glycoprotein of boar spermatozoa. FEBS Lett 280: 183–186CrossRefGoogle Scholar
  34. Jones R (1991) Interaction of zona pellucida glycoproteins, sulfated carbohydrates and synthetic polymers with proacrosin, the putative egg-binding protein from mammalian spermatozoa. Development 111: 1155–1163PubMedGoogle Scholar
  35. Jones R, Brown CR (1987) Identification of a zona-binding protein from boar spermatozoa as proacrosin. Exp Cell Res 171: 503–508PubMedCrossRefGoogle Scholar
  36. Jones R, Williams RM (1990) Identification of zona-and fucoidan-binding protein in guinea-pig spermatozoa and mechanisms of recognition. Development 109: 41–50PubMedGoogle Scholar
  37. Jones R, Brown CR, Lancaster T (1988) Carbohydrate-binding properties of boar sperm proacrosin and assessment of its role in sperm-egg recognition and adhesion during fertilization. Development 102: 781–792Google Scholar
  38. Kinloch RA, Roller RJ, Fimiani CM, Wassarman DA, Wassarman PM (1988) Primary structure of the mouse sperm receptor polypeptide determined by genomic cloning. Proc Natl Acad Sci USA 85: 6409–6413PubMedCrossRefGoogle Scholar
  39. Kinloch RA, Ruiz-Seiler B, Wassarman PM (1990) Genomic organization and polypeptide primary structure of zona pellucida glycoprotein hZP3, the hamster sperm receptor. Dev Biol 142: 414–421PubMedCrossRefGoogle Scholar
  40. Kornblihtt AR, Umezawa K, Vibe-Pedersen K, Baralle FE (1985) Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J 4: 1755–1759PubMedGoogle Scholar
  41. Leyton L, Saling P (1989a) 95kDA sperm proteins bind ZP3 and serve as tyro- sine kinase substrates in response to zona binding. Cell 57: 1123–1130Google Scholar
  42. Leyton L, Saling P (1989b) Evidence that aggregation of mouse sperm recep- tors by ZP3 triggers the acrosome reaction. J Cell Biol 108: 2163–2168PubMedCrossRefGoogle Scholar
  43. Leyton L, Robinson A, Saling P (1989) Relationship between the M42 antigen of mous sperm and the acrosome reaction induced by ZP3. Dev Biol 132: 174–178PubMedCrossRefGoogle Scholar
  44. Lillie FR (1913) The mechanism of fertilization. Science 38: 524–528PubMedCrossRefGoogle Scholar
  45. Macek MB, Shur BD (1988) Protein-carbohydrate complementarity in mammalian gamete recognition. Gamete Res 20: 93–109PubMedCrossRefGoogle Scholar
  46. Macek MB, Lopez L, Shur BD (1991) Aggregation of ß-1,4-galactosyltransferase on mouse sperm induces the acrosome reaction. Dev Biol 147: 440444Google Scholar
  47. Miller DJ, Ax RL (1990) Carbohydrates and fertilization in animals. Mol Re-prod Develop 26: 184–198CrossRefGoogle Scholar
  48. Miller DJ, Winer MA, Ax RL (1990) Heparin-binding proteins from seminal plasma bind to bovine spermatozoa and modulate capacitation by heparin. Biol Reprod 42: 899–915PubMedCrossRefGoogle Scholar
  49. Mortillo S, Wassarman PM (1991) Differential binding of gold-labeled zona pellucida glycoproteins mZP2 and mZP3 to mouse sperm membrane compartments. Development 112: 141–149Google Scholar
  50. O’Rand MG (1988) Sperm-egg recognition and barriers to interspecies fertilization. Gamete Res 19: 315–328PubMedCrossRefGoogle Scholar
  51. O’Rand MG, Widgren EE (1989) Molecular biology of a sperm antigen: Identification of the sequence of an autoantigenic epitope. In: Mettler L, Billing-ton WD (eds) Reproductive immunology. Elsevier, Amsterdam, pp 61–67Google Scholar
  52. O’Rand MG, Matthews JE, Welch JE, Fisher SJ (1985) Identification of zona binding proteins of rabbit, pig, human and mouse spermatozoa on nitrocellulose blots J Exp Zool 235: 423–428Google Scholar
  53. O’Rand MG, Welch JE, Fisher SJ (1986) Sperm membrane and zona pellucida interactions during fertilization. In: Dhindsa DS, Bahl OP (eds) Molecular and cellular aspects of reproduction. Plenum Press, New York, pp 131–144CrossRefGoogle Scholar
  54. O’Rand MG, Widgren EE, Fisher SJ (1988) Characterization of the rabbit sperm membrane autoantigen, RSA, as alectin-like zona binding protein. Develop Biol 129: 231–240PubMedCrossRefGoogle Scholar
  55. Peterson RN, Russell LD, Bundman D, Conway M, Freund M (1981) The interaction of living boar sperm and sperm plasma membrane vesicles with the porcine zona pellucida. Dev Biol 84: 144–156PubMedCrossRefGoogle Scholar
  56. Peterson RN, Russell LD, Hunt WP (1984) Evidence for specific binding of uncapacitated boar spermatozoa to porcine zonae pellucida in vitro. J Exp Zool 231: 137–147PubMedCrossRefGoogle Scholar
  57. Philpott CC, Ringuette MJ, Dean J (1987) Oocyte-specific expression and developmental regulation of ZP3, the sperm receptor of the mouse zona pellucida. Dev biol 121: 568–575PubMedCrossRefGoogle Scholar
  58. Poirier GR, Robinson R, Richardson R, Hinds K, Clayton D (1986) Evidence for a binding site on the sperm plasma membrane which recognizes the murine zona pellucida: a binding site on the sperm plasma membrane. Gamete Res 14: 235–243CrossRefGoogle Scholar
  59. Quiocho FA (1988) Molecular features and basic understanding of protein-carbohydrate interactions: the arabinose-binding protein-sugar complex. Curr Top Microbiol Immunol 139: 135–148PubMedGoogle Scholar
  60. Ringuette MJ, Sobieski DA, Chamow SM, Dean J (1986) Oocyte-specific gene expression: Molecular characterization of a cDNA coding for ZP-3, the sperm receptor of the mouse zona pellucida. Proc Natl Acad Sci USA 83: 4341–4345PubMedCrossRefGoogle Scholar
  61. Ringuette MJ, Chamberlain ME, Baur AQ, Sobieski DA, Dean J (1988) Molecular analysis of cDNA coding for ZP3, a sperm binding protein of the mouse zona pellucida. Dev Biol 127: 287–295PubMedCrossRefGoogle Scholar
  62. Robinson R, Richardson R, Hinds K, Clayton D, Poirier GR (1987) Features of a seminal proteinase inhibitor-zona pellucida-binding component on murine spermatozoa. Gamete Res 16: 217–228PubMedCrossRefGoogle Scholar
  63. Rogers BJ, Bentwood BJ (1982) Capacitation, acrosome reaction, and fertilization. In: Zaneveld LJD, Chatterton RT (eds) Biochemistry of mammalian reproduction. J. Wiley and Sons, New York, pp 203–230Google Scholar
  64. Sanz L, Calvete JJ, Mann K, Schäfer W, Schmid ER, Töpfer-Petersen E (1991) The complete primary structure of AQN-3, a carbohydrate-binding protein isolated from boar sperm. Location of disulphide bridges. FEBS Lett 291: 33–36PubMedCrossRefGoogle Scholar
  65. Sanz L, Calvete JJ, Schäfer W, Mann K, Töpfer-Petersen (1992a) Isolation and biochemical characterization of two isoforms of a boar sperm zona pellucida-binding protein. Biochem Biophys Acta (in press)Google Scholar
  66. Sanz L, Calvete JJ, Schäfer W, Schmid ER, Töpfer-Petersen E (1992b) The complete primary structure of the boar spermadhesin AQN-1, a carbohydrate-binding protein involved in fertilization. Eur J Biochem (in press)Google Scholar
  67. Sanz L, Calvete JJ, Mann K, Schäfer W, Schmid ER, Amselgruber W, Sinowatz F, Ehrhard M, Töpfer-Petersen E (1992c) The complete primary structure of the spermadhesin AWN, a zona pellucida-binding protein isolated from boar spermatozoa. FEBS Lett (in press)Google Scholar
  68. Sanz L, Calvete JJ, Jonâkovâ V, Töpfer-Petersen E (1992d) Boar spermadhesins AQN-1 and AWN are sperm-associated acrosin inhibitor acceptor proteins. FEBS Lett (in press)Google Scholar
  69. Sanz L, Calvete JJ, Mann K, Gabius H-J, Töpfer-Petersen E (1992e) Isolation and biochemical characterization of heparin-binding proteins from boar seminal plasma. A dual role for spermadhesins in fertilization. Molec Re-prod Develop (submitted)Google Scholar
  70. Schill W-B, Heimburger H, Schiesser H, Stolla R, Fritz H (1975) Reversible attachment and localization of the acid-stable seminal plasma acrosin-trypsin inhibitors on boar sperm as revealed by the indirect immunofluorescent staining technique. Hoppe-Seyler’s Z Physiol Chem 356: 1473–1476PubMedCrossRefGoogle Scholar
  71. Shur BD (1986) The receptor function of galactosyltransferase during mammalian fertilization. In: Hedrick JL (ed) The molecular and cellular biology of fertilization, Plenum Press, New York, pp 79–93CrossRefGoogle Scholar
  72. Shur BD (1988) Plasma membrane association, purification and characterization of mouse sperm ß-1,4-galactosyltransferase. J Biol Chem 263: 17706–17714PubMedGoogle Scholar
  73. Srivastava PN, Adams CE, Hartree EF (1965) Enzymatic action of acrosomal preparations on the rabbit ovum in vitro. J Reprod Fertil 10: 61–68PubMedCrossRefGoogle Scholar
  74. Töpfer-Petersen E, Henschen A (1987) Acrosin shows zona and fucose binding, novel properties for a serine proteinase. FEBS Lett 226: 38–42PubMedCrossRefGoogle Scholar
  75. Töpfer-Petersen E, Henschen A (1988) Zona pellucida-binding and fucosebinding to boar sperm acrosin is not correlated with proteolytic activity. Biol Chem Hoppe-Seyler 369: 69–76PubMedCrossRefGoogle Scholar
  76. Töpfer-Petersen E, Friess AE, Nguyen H, Schill W-B (1985) Evidence for a fucose-binding protein in boar spermatozoa. Histochem 83: 139–145CrossRefGoogle Scholar
  77. Töpfer-Petersen E, Friess AE, Henschen A, Cechova D, Steinberger M (1990a) Sperm acrosin and binding to the zona pellucida. In: Gamete interaction: prospects for immunocontraception. Wiley-Liss, Inc New York, pp 197–212Google Scholar
  78. Töpfer-Petersen E, Calvete JJ, Schäfer W, Henschen A (1990 b) Complete localization of the disulphide bridges and glycosylation sites in boar sperm acrosin. FEBS Lett 275: 139–142Google Scholar
  79. Töpfer-Petersen E, Steinberger M, Ebner von Eschenbach C, Zucker A (1990c) Zona pellucida-binding of boar sperm acrosin is associated with the N-terminal peptide of the acrosin B-chain (heavy chain). FEBS Lett 265: 51–54PubMedCrossRefGoogle Scholar
  80. Tulsiani DRP, Skudlarek MD, Orgebin-Crist MC (1989) Novel alpha-D-mannosidase of rat sperm plasma membranes: characterization and potential role in sperm-egg interactions. J Cell Biol 109: 1257–1267PubMedCrossRefGoogle Scholar
  81. Urch UA (1991) Biochemistry and function of acrosin. In: Wassarman PM (ed) Elements of mammalian fertilization, vol. 1, Basic concepts. CRC Press, Boca Raton, Florida, pp 233–248Google Scholar
  82. Urch UA, Patel H (1991) The interaction of boar sperm proacrosin with its natural substrate, the zona pellucida, and with polysulfated polysaccharides. Development 111: 1165–1172PubMedGoogle Scholar
  83. Vaquier VD, Moy GW (197) Isolation of bindin: the protein responsible for adhesion of sperm to sea urchin eggs. Proc Natl Acad Sci USA 74: 2456–2460Google Scholar
  84. Wassarman PM (1988) Zona pellucida glycoproteins. Ann Rev Biochem 57: 415–442PubMedCrossRefGoogle Scholar
  85. Wassarman PM (1990) Profile of a mammalian sperm receptor. Development 108: 1–17PubMedGoogle Scholar
  86. Yanagimachi R (1988) Mammalian fertilization. In: Knobil E, Neill J (eds) The physiologiy of reproduction. Raven Press, New York, pp 135–185Google Scholar
  87. Zaneveld LJD (1976) Sperm enzyme inhibitors as antifertility agents. In: Hafez ESE (ed) Human semen and fertility regulation in men. CV Mosby, St. Louis, pp 507–582Google Scholar
  88. Zelezna B, Cechova D (1982) Boar acrosin: isolation of two active forms of boar ejaculated sperm. Hoppe-Seyler’s Z Physiol Chem 362: 757–766CrossRefGoogle Scholar
  89. Zelezna B, Cechova D, Henschen A (1989) Isolation of the boar sperm acrosin peptide released during the conversion of a-form into 13-form. Biol Chem Hoppe-Seyler 370: 323–327PubMedCrossRefGoogle Scholar

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© Springer-Verlag Berlin Heidelberg 1992

Authors and Affiliations

  • Juan José Calvete
  • Libia Sanz
  • Edda Töpfer-Petersen

There are no affiliations available

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