Characterization of a Mouse ZP3-Derived Glycopeptide, gp55, That Exhibits Sperm Receptor and Acrosome Reaction-Inducing Activity<i>in Vitro</i>

Litscher, Eveline S.; Wassarman, Paul M.

doi:10.1021/bi952722m

Cited by 67 publications

(61 citation statements)

References 39 publications

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“…The first type is intimately bonded to the egg plasma membrane and is most extensively characterized in the sea urchin (14,15,26). The second type of egg envelope is elevated from the egg plasma membrane and is found in mollusks (7,8,27), ascidians (28)(29)(30), amphibians (31,32), and mammals (9,33,34). This discussion will pertain only to envelopes that are elevated from the plasma membrane and that the sperm must penetrate to reach the egg plasma membrane.…”

Section: Discussionmentioning

confidence: 99%

The abalone egg vitelline envelope receptor for sperm lysin is a giant multivalent molecule

Swanson

Vacquier

1997

Proc. Natl. Acad. Sci. U.S.A.

114

149

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Abalone sperm lysin is a 16-kDa acrosomal protein, which nonenzymatically and species selectively creates a hole in the egg vitelline envelope (VE) through which the sperm passes to reach the egg cell membrane. The crystal structures of both monomeric and dimeric lysins have been solved and the sequences of lysins from 20 abalone species have been determined. As a first step in understanding the molecular mechanism by which lysin creates a hole in the VE, its VE receptor was isolated. The VE receptor for lysin (VERL) is an unbranched, rod-like molecule with an approximate relative molecular mass of 2 million; half the mass being carbohydrate. Fluorescence polarization studies showed positive cooperativity in the binding of lysin to VERL (EC 50 Ϸ9 nM) and were consistent with the species selectivity of lysin in dissolving VEs. Each molecule of VERL bound between 126 and 142 molecules of monomeric lysin (two independent assays), showing that VERL possesses repetitive lysin-binding motifs.Many marine invertebrate species spawn gametes into seawater, where fertilization and embryogenesis occur. In sea urchins (1, 2) and abalone (3, 4), several closely related, congeneric species may have overlapping breeding seasons and habitats. Such species usually exhibit species selectivity (specificity) in experimental cross-fertilizations, meaning that fertilization occurs more readily between homospecific mixtures of sperm and eggs than between heterospecific mixtures. The demonstrated species selectivity in the recognition events between invertebrate sperm and egg is a valuable model bearing on the general problem of the biochemistry of molecular recognition between cells.In marine invertebrates, the study of gamete recognition proteins may also yield information on the mechanism by which species arise in the sea, where barriers to larval transport are not readily apparent (5). Evidence that gamete recognition proteins may be involved in speciation comes from analyses of nucleotide substitutions in sea urchin sperm bindins (1) and abalone sperm acrosomal proteins (4, 6). Such analyses show that the evolution of these proteins has been promoted by positive Darwinian selection, suggesting there is significant adaptive value in altering their primary structure.The availability of large quantities of marine invertebrate gametes permits a detailed characterization of their gamete surface molecules involved in fertilization. The abalone egg is surrounded by an elevated, glycoproteinaceous vitelline envelope (VE) 0.6 m in diameter (7,8). Spermatozoa bind to the VE, their acrosomal granule exocytoses, and lysin is released. Lysin creates a 3-m diameter hole in the VE through which the spermatozoan passes to reach the egg cell membrane. Electron micrographs of the lysin-created hole show it is composed of splayed-apart fibers of 13-nm diameter; covalent bonds are not broken when lysin disrupts the VE (7).Although gamete recognition proteins have been cloned from both mammals (9-13) and invertebrates (1-4, 6, 14-16), the molec...

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Section: Discussionmentioning

confidence: 99%

The abalone egg vitelline envelope receptor for sperm lysin is a giant multivalent molecule

Swanson

Vacquier

1997

Proc. Natl. Acad. Sci. U.S.A.

114

149

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“…Overall, the primary structures of ZP3 proteins are highly conserved in fish species as revealed by sequence analysis. It has been reported that the O-linked oligosaccharides of ZP3 play an important role in the recognition and adhesion of sperm and eggs (Kinloch et al, 1995;Litscher and Wassarman, 1996). Additionally, the number of O-linked glycosylation sites in the ZP3 proteins seemed to vary between different fish species.…”

Section: Zp3 In Triploid Carassius Auratus Var Pingxiangnensismentioning

confidence: 99%

Expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of ZP3 protein in oocytes in a natural, wild triploid crucian carp mutant, Carassius auratus var. Pingxiangnensis

Shi¹,

Sheng²,

Peng³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. Carassius auratus var. Pingxiangnensis (designated CaP), distributed in the Pingxiang region of Jiangxi Province, China, is a natural, wild triploid crucian carp mutant that has two reproductive development modes: gynogenesis and bisexual reproduction. Little information is available about the expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of the ZP3 protein in oocytes of this fish. In this study, we obtained the full-length cDNA of ZP3 (CaP_ ZP3). CaP_ZP3 contains an open reading frame of 1305 bp that encodes 435 amino acid residues. Real-time polymerase chain reaction (PCR) was used to determine the CaP_ZP3 mRNA expression levels in the ovary at different stages of maturation. Results revealed high levels of CaP_ZP3 expression in 4-to 8-month-old ovaries (stage II-stage III), with a significant decline in 9-to 12-month-old ovaries (stages IV-stage V). The high levels of CaP_ ZP3 transcripts during the early growth period suggest an important role for CaP_ZP3 in early oocyte development. In addition, a polyclonal antibody was prepared against CaP_ZP3, and the immunofluorescence localization was determined. CaP_ZP3 protein was detected close to the oocyte plasma membrane. The results also showed that no fluorescent signal was detected in stage I and II oocytes. CaP_ZP3 protein is primarily detected in stage III oocytes, and the protein accumulates as oocytes develop into stage IV oocytes. These results suggested that the transcription and translation of the CaP_ZP3 gene is asynchronous and that the transcription of the CaP_ZP3 protein occurs prior to its translation in this triploid fish.

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“…Several reports have concluded that the C-terminal region of mZP3 polypeptide is responsible for the ability of mZP3 to inhibit binding of sperm to eggs and to induce the AR (Rosiere and Wassarman, 1992;Kinloch et al, 1995;Litscher and Wassarman, 1996b;Li et al, 2007). In this context, Millar et al (1989) reported that a short peptide (16 amino acids) from the C-terminal region of mZP3 produced antibodies in mice to mZP3.…”

Section: Relevant Observations and Implicationsmentioning

confidence: 99%

Zona pellucida glycoprotein ZP3 and fertilization in mammals

Litscher

Williams

Wassarman

2009

Molecular Reproduction Devel

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SUMMARYAn early step in mammalian fertilization is species-restricted binding of sperm to the egg's zona pellucida (ZP), a thick extracellular coat that surrounds eggs. Sperm bind to the ZP of unfertilized eggs, but not to the ZP of fertilized eggs. Shortly after binding to the unfertilized egg ZP, sperm undergo the acrosome reaction, a form of cellular exocytosis that enables sperm to penetrate the ZP. Three glycoproteins, mZP1-3, constitute the mouse egg's ZP and participate in the process of fertilization. For example, sperm exposed to unfertilized egg mZP3 at nanomolar concentrations are inhibited from binding to eggs and undergo the acrosome reaction. Neither mZP1 nor mZP2 has an effect on sperm binding or the acrosome reaction. Furthermore, mZP3 from fertilized eggs has no effect on sperm binding and is unable to induce the acrosome reaction. These and other properties of mZP3 suggest that it is a receptor for sperm and inducer of the acrosome reaction. Mapping of the mZP3 combining-site for sperm suggests that it is located near the C-terminus of the polypeptide, just downstream of the ZP domain, in a region encoded by exon-7 of the mZP3 gene. This region of mZP3 is a site of positive Darwinian selection. When mZP3 exon-7 is fused to the Fc fragment of human IgG and sperm exposed to the chimeric protein, sperm are inhibited from binding to eggs. However, the chimeric protein does not induce the acrosome reaction. Therefore, polypeptide encoded by mZP3 exon-7 is necessary and sufficient for binding of mouse sperm.

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Characterization of a Mouse ZP3-Derived Glycopeptide, gp55, That Exhibits Sperm Receptor and Acrosome Reaction-Inducing Activityin Vitro

Cited by 67 publications

References 39 publications

The abalone egg vitelline envelope receptor for sperm lysin is a giant multivalent molecule

The abalone egg vitelline envelope receptor for sperm lysin is a giant multivalent molecule

Expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of ZP3 protein in oocytes in a natural, wild triploid crucian carp mutant, Carassius auratus var. Pingxiangnensis

Zona pellucida glycoprotein ZP3 and fertilization in mammals

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