Lewis X-containing neoglycoproteins mimic the intrinsic ability of zona pellucida glycoprotein ZP3 to induce the acrosome reaction in capacitated mouse sperm

William F. Hanna, Candace L. Kerr, Joel H. Shaper, William W Wright

Research output: Contribution to journalArticle

Abstract

The binding of zona pellucida (ZP) glycoprotein ZP3 to mouse sperm surface receptors is mediated by protein-carbohydrate interactions. Subsequently, ZP3 induces sperm to undergo the acrosome reaction, an obligatory step in fertilization. We have previously identified Lewis X (Lex; Galβ4[Fucα3] GlcNAc) as a potent inhibitor of in vitro sperm-ZP binding (Johnston et al. J Biol Chem 1998; 273:1888-1895). This glycan is recognized by ∼70% of the ZP3 binding sites on capacitated, acrosome-intact mouse sperm, whereas Lewis A (Lea; Galβ3[Fucα4]GlcNAc) is recognized by most of the remaining sites (Kerr et al. Biol Reprod 2004; 71:770-777). Herein, we test the hypothesis that Lex- and Le a-containing glycans, when clustered on a neoglycoprotein, bind ZP3 receptors on sperm and induce sperm to undergo the acrosome reaction via the same signaling pathways as ZP3. Results show that a Lex-containing neoglycoprotein induced the acrosome reaction in a dose-dependent and capacitation-dependent manner. A Lea-containing neoglycoprotein also induced sperm to undergo the acrosome reaction but was less potent than Le x-containing neoglycoproteins. In contrast, neoglycoproteins containing β4-lactosamine (Galβ4GlcNAc), Lewis B (Fucα2Galβ 3[Fucα4]GlcNAc), and sialyl-Lex glycans were inactive, as were four other neoglycoproteins with different nonfucosylated glycans. Consistent with these results, unconjugated Lex- and Lea-capped glycans were dose-dependent inhibitors, which at saturation, reduced the ZP-induced acrosome reaction by about 60% and 30%, respectively. Experiments utilizing pharmacological inhibitors suggest that induction of the acrosome reaction by solubilized ZP and Lex- and Lea-containing neoglycoproteins require the same calcium-dependent pathway. However, only the ZP-induced acrosome reaction requires a functional Gi protein. Thus, Lex-containing neoglycoproteins bind to a major class of ZP3 receptors on capacitated sperm. A Lea-containing neoglycoprotein binds a second ZP3 receptor but is a less-potent inducer of the acrosome reaction.

Original languageEnglish (US)
Pages (from-to)778-789
Number of pages12
JournalBiology of Reproduction
Volume71
Issue number3
DOIs
StatePublished - Sep 2004

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Acrosome Reaction
Spermatozoa
Glycoproteins
Zona Pellucida
Polysaccharides
Zona Pellucida Glycoproteins
Acrosome
Fertilization
Proteins
Binding Sites
Carbohydrates
Pharmacology
Calcium

Keywords

  • Acrosome reaction
  • Fertilization
  • Gamete biology
  • Sperm

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Lewis X-containing neoglycoproteins mimic the intrinsic ability of zona pellucida glycoprotein ZP3 to induce the acrosome reaction in capacitated mouse sperm. / Hanna, William F.; Kerr, Candace L.; Shaper, Joel H.; Wright, William W.

In: Biology of Reproduction, Vol. 71, No. 3, 09.2004, p. 778-789.

Research output: Contribution to journalArticle

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abstract = "The binding of zona pellucida (ZP) glycoprotein ZP3 to mouse sperm surface receptors is mediated by protein-carbohydrate interactions. Subsequently, ZP3 induces sperm to undergo the acrosome reaction, an obligatory step in fertilization. We have previously identified Lewis X (Lex; Galβ4[Fucα3] GlcNAc) as a potent inhibitor of in vitro sperm-ZP binding (Johnston et al. J Biol Chem 1998; 273:1888-1895). This glycan is recognized by ∼70{\%} of the ZP3 binding sites on capacitated, acrosome-intact mouse sperm, whereas Lewis A (Lea; Galβ3[Fucα4]GlcNAc) is recognized by most of the remaining sites (Kerr et al. Biol Reprod 2004; 71:770-777). Herein, we test the hypothesis that Lex- and Le a-containing glycans, when clustered on a neoglycoprotein, bind ZP3 receptors on sperm and induce sperm to undergo the acrosome reaction via the same signaling pathways as ZP3. Results show that a Lex-containing neoglycoprotein induced the acrosome reaction in a dose-dependent and capacitation-dependent manner. A Lea-containing neoglycoprotein also induced sperm to undergo the acrosome reaction but was less potent than Le x-containing neoglycoproteins. In contrast, neoglycoproteins containing β4-lactosamine (Galβ4GlcNAc), Lewis B (Fucα2Galβ 3[Fucα4]GlcNAc), and sialyl-Lex glycans were inactive, as were four other neoglycoproteins with different nonfucosylated glycans. Consistent with these results, unconjugated Lex- and Lea-capped glycans were dose-dependent inhibitors, which at saturation, reduced the ZP-induced acrosome reaction by about 60{\%} and 30{\%}, respectively. Experiments utilizing pharmacological inhibitors suggest that induction of the acrosome reaction by solubilized ZP and Lex- and Lea-containing neoglycoproteins require the same calcium-dependent pathway. However, only the ZP-induced acrosome reaction requires a functional Gi protein. Thus, Lex-containing neoglycoproteins bind to a major class of ZP3 receptors on capacitated sperm. A Lea-containing neoglycoprotein binds a second ZP3 receptor but is a less-potent inducer of the acrosome reaction.",
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