Affinage

ZP3

Zona pellucida sperm-binding protein 3 · UniProt P21754

Length
424 aa
Mass
47.0 kDa
Annotated
2026-04-28
100 papers in source corpus 34 papers cited in narrative 35 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ZP3 is an oocyte-secreted glycoprotein that serves as the essential structural scaffold of the zona pellucida extracellular matrix and as the primary sperm receptor mediating species-restricted gamete recognition. ZP3 is synthesized with a signal peptide and trafficked independently of ZP2 through the oocyte endomembrane system; its cytoplasmic tail prevents premature intracellular ZP2–ZP3 interaction, and secretion requires furin-site cleavage near the C-terminus and an intact conserved hydrophobic patch (EHP) that also prevents premature polymerization (PMID:11170414, PMID:14645511, PMID:21378311). Once incorporated into the zona pellucida via ZP domain–mediated polymerization—whose structural basis was revealed by crystallography of the ZP-N immunoglobulin-like fold—ZP3 presents O-linked oligosaccharides on its exon-7-encoded combining site that bind sperm surface proteins (sp56/ZP3R, GalTase), triggering receptor aggregation and a multi-branch signaling cascade involving pertussis toxin–sensitive G proteins, phospholipase C, PI3K/Akt/PKCζ, T-type Ca²⁺ channel activation, and Trp2-dependent sustained store-operated Ca²⁺ entry that drives the acrosome reaction (PMID:2986849, PMID:7816829, PMID:19052627, PMID:2544604, PMID:11331878, PMID:17258189). Homozygous Zp3 disruption in mice abolishes zona pellucida formation and fertility, and dominant-negative ZP3 mutations in humans (e.g., p.Ala134Thr) cause empty follicle syndrome by blocking the ZP3–ZP2 interaction required for matrix assembly (PMID:8787763, PMID:28886344).

Mechanistic history

Synthesis pass · year-by-year structured walk · 16 steps
  1. 1985 High

    Establishing which glycan class on ZP3 mediates sperm binding resolved whether carbohydrates or protein mediate gamete recognition, showing that O-linked—not N-linked—oligosaccharides carry the sperm receptor activity.

    Evidence Enzymatic removal of O-linked vs N-linked oligosaccharides from purified ZP3 followed by sperm binding assays in mouse

    PMID:2986849

    Open questions at the time
    • Identity of the specific O-glycan structure(s) mediating binding was unknown
    • Sperm-side receptor for these O-glycans not yet identified
  2. 1988 High

    Defining the primary structure of ZP3 and the terminal sugar requirement for sperm binding established the molecular framework—a secreted glycoprotein whose alpha-linked galactose residues on O-glycans are critical for receptor function.

    Evidence cDNA cloning of mouse ZP3 revealing signal peptide, glycosylation sites, and conserved sequences; enzymatic/chemical modification showing alpha-galactose requirement

    PMID:2842789 PMID:3378665

    Open questions at the time
    • Whether alpha-1,3-galactose specifically is required was unresolved
    • The polypeptide region presenting these glycans was unknown
  3. 1989 High

    Demonstrating that ZP3-induced receptor aggregation on sperm is sufficient to trigger the acrosome reaction, and identifying a 95 kDa tyrosine kinase substrate as a candidate ZP3 receptor, established the signaling logic: multivalent ZP3 cross-links sperm receptors to initiate exocytosis.

    Evidence Anti-ZP3 IgG vs Fab aggregation assays with acrosome reaction readout; photoaffinity crosslinking and phosphotyrosine immunoblotting on sperm

    PMID:2472220 PMID:2544604

    Open questions at the time
    • Molecular identity of the 95 kDa kinase substrate was uncertain
    • Downstream signaling intermediates were unknown
  4. 1990 High

    Identification and localization of sp56 as a ZP3-binding protein on acrosome-intact sperm heads provided a candidate sperm-side receptor that could explain the specificity of ZP3 recognition.

    Evidence Photoaffinity crosslinking, ZP3 affinity chromatography, and immunolocalization on mouse sperm

    PMID:2371290 PMID:8188752

    Open questions at the time
    • Genetic requirement of sp56 for fertilization not yet demonstrated
    • Relationship of sp56 to other proposed ZP3-binding proteins (GalTase, 95 kDa) unresolved
  5. 1992 High

    Discovery of the ZP domain as a shared ~260-residue module, and demonstration that sperm GalTase binds specifically to ZP3 O-glycans whose modification post-fertilization abolishes binding, connected ZP3 structure to the polyspermy block mechanism.

    Evidence Sequence analysis identifying ZP domain across multiple proteins; competitive binding and enzymatic modification of ZP3 with cortical granule modification experiments

    PMID:1313375 PMID:1608469

    Open questions at the time
    • Three-dimensional structure of ZP domain unknown
    • How cortical granule enzymes specifically modify ZP3 O-glycans uncharacterized
  6. 1995 High

    Exon-swapping and mutagenesis pinpointed exon 7 as encoding the sperm combining site, while knockout of alpha-1,3-galactosyltransferase showed the Gal-alpha-1,3-Gal epitope is dispensable, refining the glycan specificity model.

    Evidence Site-directed mutagenesis of serine glycosylation sites in EC cells with sperm binding assays; alpha-1,3GT knockout mice with fertility assessment

    PMID:7545161 PMID:7816829

    Open questions at the time
    • Exact O-glycan structures at specific sites on native ZP3 not yet determined by mass spectrometry
    • Whether species specificity maps entirely to exon 7 sequence divergence was unresolved
  7. 1996 High

    ZP3 knockout mice lacking any zona pellucida demonstrated that ZP3 is the essential nucleating component for matrix assembly and is required for female fertility, establishing its dual structural and signaling roles.

    Evidence Homologous recombination Zp3 knockout in mouse with immunofluorescence, electron microscopy, and fertility testing

    PMID:8787763

    Open questions at the time
    • Whether ZP3 initiates filament nucleation or merely co-assembles with ZP1/ZP2 was mechanistically unresolved
    • Contribution of ZP3 to oocyte maturation beyond zona assembly not explored
  8. 1996 High

    Defining the sperm signaling cascade downstream of ZP3—membrane depolarization via a cation channel, pertussis toxin–sensitive G-protein activation, and voltage-sensitive Ca²⁺ channel opening—placed ZP3 at the head of a multi-branch signal transduction pathway driving the acrosome reaction.

    Evidence Potentiometric fluorescent probes and pharmacological dissection in mouse sperm

    PMID:8707844

    Open questions at the time
    • Identity of the ZP3-activated cation channel unknown
    • Sustained Ca²⁺ entry mechanism not yet characterized
  9. 2001 High

    Identification of Trp2 (TRPC2) as the channel mediating ZP3-triggered sustained store-operated Ca²⁺ entry, and of furin-site cleavage as essential for ZP3 secretion, resolved both the terminal Ca²⁺ entry step in acrosome reaction signaling and a key post-translational processing requirement.

    Evidence Trp2 genetic analysis with Ca²⁺ imaging and pharmacology; furin-site mutagenesis and inhibitor studies in transfected cells

    PMID:11170414 PMID:11331878

    Open questions at the time
    • How store depletion activates Trp2 in sperm was unresolved
    • Whether furin or another proprotein convertase performs the cleavage in oocytes in vivo was not determined
  10. 2003 High

    Linker-scanning mutagenesis in transgenic mice revealed that a conserved eight-residue hydrophobic patch (EHP) between the furin site and transmembrane domain is required for ZP3 secretion and zona incorporation, distinguishing EHP-dependent processing from furin cleavage.

    Evidence Mutagenesis of EGFP-ZP3 fusions, confocal imaging in oocytes, transgenic mice, and mass spectrometry of native ectodomain cleavage site

    PMID:14645511

    Open questions at the time
    • Mechanism by which EHP controls secretion vs polymerization was structurally unresolved
    • Whether EHP acts as an intramolecular chaperone or sorting signal was unknown
  11. 2006 High

    Mass spectrometric mapping of native ZP3 O-glycosylation sites (Thr-155 and residues 161–168) and demonstration that ZP3 activates PI3K/Akt/PKCζ signaling in sperm linked specific glycan structures to downstream fertilization-essential kinase cascades.

    Evidence MS O-glycan analysis of zona proteins from transgenic mice; lipid kinase assays and PI3K inhibitors with IVF readout

    PMID:16401092 PMID:17258189

    Open questions at the time
    • Which specific O-glycan structure at these sites is the functional sperm ligand remained debated
    • How PI3K pathway intersects with G-protein and Ca²⁺ signaling branches was not mapped
  12. 2008 High

    The 2.3 Å crystal structure of the ZP3 ZP-N domain revealed an immunoglobulin-like fold with a polymerization-implicated E' strand and invariant tyrosine, providing the first atomic-level understanding of zona pellucida filament assembly.

    Evidence X-ray crystallography of mouse ZP3 ZP-N domain

    PMID:19052627

    Open questions at the time
    • Full ZP domain structure including ZP-C half was not yet resolved
    • How ZP-N mediates heteromeric ZP2–ZP3 polymerization was structurally undetermined
  13. 2010 High

    The 2.0 Å structure of avian ZP3 showed the complete ZP domain in a homodimer, revealed how the EHP propeptide prevents premature polymerization, and identified close positioning of the O-glycan and hypervariable loop as a structural basis for species-restricted recognition.

    Evidence X-ray crystallography of chicken ZP3 homolog at 2.0 Å

    PMID:20970175

    Open questions at the time
    • Whether the homodimeric arrangement is conserved in mammalian ZP3 was unverified
    • Structural basis of intramolecular signaling upon sperm binding remained speculative
  14. 2011 High

    Demonstrating that ZP3's cytoplasmic tail prevents premature intracellular ZP2–ZP3 interaction explained how zona proteins traffic independently and polymerize only after release into the extracellular space.

    Evidence Bimolecular fluorescence complementation in CHO cells and oocytes; tail-deletion and tail-swap experiments in transgenic mice

    PMID:21378311

    Open questions at the time
    • Molecular mechanism by which the cytoplasmic tail masks the interaction interface was not determined
    • Whether other quality-control checkpoints exist in the ER was not explored
  15. 2017 High

    A dominant-negative ZP3 mutation (p.Ala134Thr) was shown to cause human empty follicle syndrome by binding wild-type ZP3 and blocking ZP3–ZP2 interaction, establishing the first direct genetic link between ZP3 mutations and human infertility.

    Evidence Patient family genetics, co-immunoprecipitation of mutant and wild-type ZP3, immunofluorescence

    PMID:28886344

    Open questions at the time
    • Whether additional ZP3 mutations cause similar phenotypes was not systematically assessed
    • Structural basis for dominant-negative interference not resolved
  16. 2020 Medium

    A second pathogenic ZP3 mutation (p.Ser173Cys) disrupting ZP3–ZP2 interaction confirmed that ZP2–ZP3 heteroassembly is the critical step for zona pellucida formation in humans.

    Evidence Whole-exome sequencing, co-immunoprecipitation, and protein localization studies

    PMID:33140178

    Open questions at the time
    • Single co-IP without reciprocal validation or structural modeling
    • Genotype-phenotype correlation across the ZP3 mutation spectrum incomplete

Open questions

Synthesis pass · forward-looking unresolved questions
  • The precise O-glycan structure on ZP3 that constitutes the minimal sperm-binding determinant remains unidentified, and the complete structural basis for species-restricted sperm recognition—including how sperm receptor binding triggers conformational changes in ZP3 that propagate into the signaling cascade—is unresolved.
  • No consensus on the specific O-glycan determinant required for sperm binding despite decades of study
  • Full-length mammalian ZP3 structure in complex with a sperm receptor has not been determined
  • How the ZP3-initiated signaling cascade integrates G-protein, PI3K, and Trp2/Ca²⁺ branches into a unified exocytotic decision remains unmapped

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0048018 receptor ligand activity 8 GO:0005198 structural molecule activity 3
Localization
GO:0005576 extracellular region 5 GO:0005886 plasma membrane 2 GO:0005783 endoplasmic reticulum 1
Pathway
R-HSA-1474165 Reproduction 7 R-HSA-1474244 Extracellular matrix organization 4 R-HSA-162582 Signal Transduction 4
Partners
B4GALT1CRISP1TRPC2ZP1ZP2ZP3R
Complex memberships
Zona pellucida matrix (ZP1–ZP2–ZP3)

Evidence

Reading pass · 35 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1985 O-linked oligosaccharides on mouse egg ZP3, but not N-linked oligosaccharides, are responsible for sperm receptor activity; a specific size class of O-linked oligosaccharides recovered from ZP3 possesses sperm receptor activity and binds sperm directly. Enzymatic removal of O-linked vs N-linked oligosaccharides from ZP3 followed by sperm binding assays Cell High 2986849
1988 Galactose in alpha-linkage at the nonreducing terminus of O-linked oligosaccharides on ZP3 is essential for sperm receptor activity; alpha-galactosidase or galactose oxidase treatment abolishes activity, which can be restored by sodium borohydride reduction. Enzymatic and chemical modification of ZP3 O-linked oligosaccharides with sperm receptor activity assays Proceedings of the National Academy of Sciences of the United States of America High 2842789
1988 Mouse ZP3 encodes a polypeptide of 46,307 Da with a 22-amino acid N-terminal signal peptide, six potential N-linked glycosylation sites, and sequences conserved among mammals; the protein is secreted after signal peptide cleavage. cDNA cloning and molecular characterization Developmental biology High 3378665
1989 A 95 kDa sperm surface protein binds ZP3 and serves as a tyrosine kinase substrate upon ZP3 binding; the same 95 kDa protein that binds ZP3 shows enhanced phosphotyrosine immunoreactivity in a ZP concentration-dependent manner, suggesting it is both a ZP3 receptor and a tyrosine kinase substrate initiating the acrosome reaction cascade. Photoaffinity crosslinking, radiolabeled ZP3 blot binding, anti-phosphotyrosine immunoblotting, immunofluorescence localization Cell High 2472220
1989 ZP3 aggregation of sperm surface receptors is sufficient to trigger the acrosome reaction; anti-ZP3 IgG (but not Fab fragments) induced acrosome reactions in sperm pre-incubated with ZP3 glycopeptides, and cross-linking Fab fragments with secondary antibody restored this activity, demonstrating receptor aggregation as the triggering mechanism. Antibody-mediated receptor aggregation assays with anti-ZP2, anti-ZP3 IgG and Fab fragments; chlortetracycline fluorescence assay for acrosome reaction The Journal of cell biology High 2544604
1990 A Mr 56,000 sperm protein (sp56) binds specifically to ZP3 via photoaffinity crosslinking, binds tightly to ZP3 affinity columns, and is localized to heads of acrosome-intact but not acrosome-reacted sperm, identifying it as a ZP3-binding protein on the sperm surface. Photoaffinity crosslinking with Denny-Jaffee reagent, ZP3 affinity chromatography, immunolocalization Proceedings of the National Academy of Sciences of the United States of America High 2371290
1991 Mouse embryonal carcinoma (EC) cells transfected with ZP3 gene produce and secrete biologically active mZP3 that retains sperm receptor and acrosome reaction-inducing activity, confirming that EC cell glycosylation machinery adds the specific O-linked oligosaccharides essential for ZP3 biological activity. Stable transfection of EC cells, purification of recombinant ZP3, sperm binding inhibition and acrosome reaction assays The Journal of cell biology High 1655811
1992 Sperm surface beta-1,4-galactosyltransferase (GalT) binds specifically to ZP3 O-linked oligosaccharides that have sperm-binding activity but not other zona pellucida glycoproteins; blocking or removing the GalT binding site on ZP3 inhibits sperm-egg binding; after acrosome reaction, GalT relocalizes so it can no longer bind ZP3; and ZP3 modification post-fertilization by cortical granule secretions removes the GalT binding site, accounting for loss of sperm receptor activity. Competitive binding assays, enzymatic modification of ZP3, immunolocalization of GalT on acrosome-intact vs. acrosome-reacted sperm, cortical granule modification experiments Nature High 1608469
1992 A ZP domain (approximately 260 residues) is shared between sperm receptors ZP2 and ZP3 and other proteins including betaglycan (TGF-beta type III receptor), uromodulin, and GP-2, suggesting common tertiary structure and functional similarity in recognition mechanisms. Sequence analysis and domain identification FEBS letters Medium 1313375
1992 Recombinant mouse ZP3 expressed in rodent and primate cells is biologically active, inhibiting sperm-zona binding and triggering acrosomal exocytosis in capacitated mouse sperm at concentrations equivalent to native ZP3. Stable transfection, recombinant protein purification, sperm binding inhibition assay, acrosome reaction assay Developmental biology High 1577197
1994 Sperm protein sp56 (Mr 56,000) purified by ZP3 affinity chromatography is a peripheral plasma membrane protein on the outer surface of the sperm head that specifically recognizes and binds the ZP3 oligosaccharide; it forms a homomultimer (~110,000 Mr) with intramolecular disulfide bonds. ZP3 affinity chromatography, ion exchange chromatography, size-exclusion chromatography, monoclonal antibody immunohistochemistry and immunoblotting, crosslinking experiments The Journal of cell biology High 8188752
1994 Recombinant human ZP3 produced in CHO cells induces the human sperm acrosome reaction and promotes sperm-egg fusion with zona-free hamster oocytes, demonstrating biological activity of the recombinant protein despite slightly lower molecular weight than native ZP3. CHO cell expression of recombinant human ZP3, Western blot, acrosome reaction assay, sperm-egg fusion assay Biology of reproduction High 7819440
1995 Site-directed mutagenesis mapping of mZP3 shows that glycosylation of serine residues clustered in the polypeptide region encoded by exon 7 is required for both sperm binding and acrosome reaction-inducing activities; exon 7 encodes the sperm combining site and exhibits sequence divergence consistent with species-specific gamete adhesion. Exon swapping, site-directed mutagenesis, stable transfection of embryonal carcinoma cells, sperm binding and acrosome reaction assays Proceedings of the National Academy of Sciences of the United States of America High 7816829
1995 The Gal alpha1,3Gal epitope displayed on ZP3 is not required for fertilization, as mice genetically deficient in the alpha1,3-galactosyltransferase enzyme and lacking this epitope on ZP3 are fully fertile. Gene knockout (alpha1,3GT-/- mice), fertility testing, biochemical verification of epitope absence The Journal of biological chemistry High 7545161
1996 Homozygous insertional mutation of Zp3 in mice results in complete absence of zona pellucida matrix despite the presence of ZP1 and ZP2 in oocytes, demonstrating ZP3 is essential for zona matrix assembly and fertility; Zp3-/- oocytes lack organized corona radiata and fail to produce 2-cell embryos after mating. Homologous recombination knockout, immunofluorescence, electron microscopy, fertility assessment Development (Cambridge, England) High 8787763
1996 ZP3 depolarizes sperm membranes by activating a pertussis toxin-insensitive cation channel, and also activates a pertussis toxin-sensitive pathway producing a transient rise in internal pH; these combined effects open voltage-sensitive Ca2+ channels to initiate the acrosome reaction. Potentiometric fluorescent probes, pharmacological inhibition with pertussis toxin, electrophysiological approaches The Journal of cell biology High 8707844
1996 A glycopeptide (gp55, ~55 kDa) derived from the carboxy-terminal half of mZP3 polypeptide exhibits both sperm receptor activity and acrosome reaction-inducing activity; N-linked oligosaccharides and sialic acid are not essential elements of the mZP3 combining site for sperm. Proteolytic fragmentation of mZP3, N-glycanase digestion, endo-beta-galactosidase treatment, neuraminidase treatment, sperm binding inhibition and acrosome reaction assays Biochemistry High 8672430
1998 Pig ZPB and ZPC form heterocomplexes that are the functional ligand for boar sperm membrane binding sites; neither ZPB nor ZPC alone binds sperm membranes at high affinity, but pZPB-pZPC heterocomplexes detected by size exclusion chromatography and Western blot show high-affinity saturable binding. Solid phase binding assays with immobilized boar sperm membranes, size exclusion chromatography, Western blot for heterocomplexes, competitive binding assays The Journal of biological chemistry High 9516448
2000 ZP3 activates a store depletion-operated Ca2+ entry pathway in sperm that sustains elevated intracellular Ca2+ concentration and drives the exocytotic acrosome reaction; ZP3 and thapsigargin activate the same Ca2+ permeation mechanism as shown by fluorescence quenching and shared channel antagonists. Fluorometric Ca2+ measurements, fluorescence quenching experiments, pharmacological antagonists, comparison with thapsigargin-activated pathway Molecular biology of the cell High 10793136
2001 Trp2 (TRPC2) channel is essential for ZP3-triggered sustained Ca2+ influx into mouse sperm; ZP3 activates trimeric G proteins and phospholipase C causing transient Ca2+ influx through T-type channels, which then activates a Trp2-dependent sustained Ca2+ entry pathway that drives acrosome reactions. Genetic analysis of Trp2, Ca2+ imaging, pharmacological dissection of early vs. late Ca2+ entry pathways Nature cell biology High 11331878
2001 Secretion of mouse ZP3 from transfected cells requires cleavage at a consensus furin cleavage-site near the C-terminus; site-directed mutagenesis of the furin site (RNRR) and use of a furin inhibitor both block ZP3 secretion, defining a novel function for furin family enzymes in ZP3 processing. Site-directed mutagenesis of furin site, furin-specific inhibitor treatment, transfection of cells, Western blot analysis of secretion Biochemistry High 11170414
2002 ZP3 and ZP2 cytoplasmic tails are necessary and sufficient to prevent intracellular oligomerization of zona proteins; in absence of cytoplasmic tails, ZP2 and ZP3 interact within the cell and fail to incorporate into the zona pellucida; ZP2 and ZP3 traffic independently to the plasma membrane where they interact only after release into the extracellular matrix. Fluorescent protein tagging, bimolecular fluorescence complementation (BiFC) in CHO cells and growing oocytes, confocal microscopy Journal of cell science High 21378311
2003 An eight-amino-acid conserved hydrophobic patch in ZP3, located between the furin cleavage site and transmembrane domain, is required for secretion and incorporation into the zona pellucida; mass spectrometry of native zona pellucida defined the precise ectodomain cleavage site N-terminal to a proprotein convertase site distinct from the hydrophobic patch. Linker-scanning mutagenesis, EGFP-ZP3 fusion protein trafficking in oocytes, confocal microscopy, transgenic mice, mass spectrometry Molecular and cellular biology High 14645511
2006 ZP2 and ZP3 traffic independently through the oocyte endomembrane system (colocalizing in ER and multivesicular aggregates without physical interaction detected by co-immunoprecipitation), and N-glycans on ZP3 are not required for its trafficking or incorporation into the zona pellucida. Co-immunoprecipitation, fluorescent protein tagging, confocal microscopy in growing oocytes, transgenic mice with N-glycosylation-deficient ZP3-EGFP Molecular and cellular biology High 17047254
2006 ZP3 stimulates production of phosphatidylinositol-(3,4,5)-triphosphate in sperm membranes; PI3-kinase antagonists prevent acrosome reactions and fertilization; downstream effectors include Akt and PKCzeta, outlining a PI3K signaling pathway essential for early fertilization events. Lipid kinase assays, pharmacological PI3K antagonists, in vitro fertilization, identification of downstream kinases Developmental biology High 17258189
2006 The mZP3 sperm combining site is encoded by exon 7; substitution of mZP3 exon-7 into hamster ZP3 imparts sperm inhibitory activity, and an IgG(Fc)/mZP3(exon-7+8) chimeric fusion protein binds specifically to acrosome-intact sperm heads and inhibits sperm-egg binding in vitro at nanomolar concentrations. Exon swapping between mouse and hamster ZP3, IgG(Fc) fusion protein construction, stable EC cell transfection, sperm binding assays Journal of cellular physiology High 16245311
2006 Two O-glycosylation sites in native mZP3 at Thr-155 and within residues 161-168 (ATVSSEEK) are conserved in human ZP3 from transgenic mice; core 2 O-glycans predominate at both sites in mZP3, while in huZP3 from rescue mice core 1 and Tn sequences predominate at the Thr-156 equivalent site; despite these differences, sperm binding is not affected. Mass spectrometry O-glycan analysis of native zona proteins from transgenic mice expressing human ZP3 Biochemistry High 16401092
2008 Crystal structure of the ZP-N domain of mouse ZP3 at 2.3 Å resolution reveals a new immunoglobulin superfamily subtype with a beta-sheet extension containing an E' strand and invariant tyrosine implicated in polymerization; the structure supports the presence of ZP-N repeats in ZP2 and other zona proteins. X-ray crystallography at 2.3 Å resolution Nature High 19052627
2008 Recombinant ZP3R/sp56 forms a high molecular weight disulfide cross-linked oligomer (six or more monomers) and binds the zona pellucida of unfertilized eggs but not 2-cell embryos; pre-incubation of eggs with recombinant ZP3R/sp56 inhibits in vitro fertilization by ~74%, demonstrating its role in sperm-zona interactions. HEK293 cell expression of recombinant ZP3R/sp56, size-exclusion chromatography, zona pellucida binding assays, in vitro fertilization inhibition assay The Journal of biological chemistry High 18316377
2010 Crystal structure of an avian ZP3 homolog at 2.0 Å resolution reveals the complete ZP domain module in a homodimeric arrangement required for secretion; an external hydrophobic patch (EHP) in the propeptide prevents premature polymerization; close positioning of a conserved O-glycan and the hypervariable C-terminal region suggests concerted role in species-restricted gamete recognition; alternative conformations around the O-glycan indicate a mechanism for downstream intramolecular signaling upon sperm binding. X-ray crystallography at 2.0 Å resolution; structural analysis of EHP and ZP domain interactions Cell High 20970175
2011 ZP3 cytoplasmic tail prevents premature intracellular interactions of ZP3 with ZP2; removal of the cytoplasmic tail allows intracellular ZP2-ZP3 interaction and prevents their incorporation into the zona pellucida matrix; the cytoplasmic tail is both necessary and sufficient for correct trafficking and zona incorporation. Bimolecular fluorescence complementation (BiFC), fluorescent tagging, confocal microscopy in CHO cells and growing oocytes, transgenic mouse analysis Journal of cell science High 21378311
2013 Human epididymal CRISP1 mediates sperm-zona pellucida binding through specific interaction with ZP3; recombinant hCRISP1 interacts with ZP3 but not ZP2 or ZP4 in dose-dependent saturable ELISA assays; inhibition of sperm hCRISP1 reduces sperm binding to the hemizona. Hemizona assay with antibody inhibition, ELISA binding assay with recombinant ZP proteins, immunofluorescence Molecular human reproduction Medium 24334245
2017 A dominant negative ZP3 mutation (p.Ala134Thr) causes empty follicle syndrome by binding to wild-type ZP3 and preventing the wild-type ZP3-ZP2 interaction required for zona pellucida assembly, thereby impeding ZP matrix formation and causing oocyte degeneration. Co-expression of wild-type and mutant ZP3, co-immunoprecipitation to detect ZP3-ZP2 interaction, immunofluorescence, patient genetics American journal of human genetics High 28886344
2017 ZP3 is required for germinal vesicle breakdown (GVBD) in mouse oocyte meiosis; ZP3 is expressed in the nucleus during prophase and translocates to the zona pellucida; ZP3 knockdown inhibits GVBD; ZP3 interacts with Ptprk, Aipl1, and Diaph2, and regulates Akt phosphorylation, lamin-nuclear membrane binding via Aipl1, and actin cytoskeleton organization via Diaph2. siRNA knockdown, immunoprecipitation with mass spectrometry to identify interacting proteins, immunofluorescence for localization Scientific reports Medium 28145526
2020 A novel ZP3 mutation (p.Ser173Cys) disrupts ZP3-ZP2 protein-protein interaction as demonstrated by co-immunoprecipitation, leading to abnormal zona pellucida formation and empty follicle syndrome. Co-immunoprecipitation, subcellular protein localization, Western blot, whole-exome sequencing Journal of assisted reproduction and genetics Medium 33140178

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1985 O-linked oligosaccharides of mouse egg ZP3 account for its sperm receptor activity. Cell 539 2986849
1992 Complementarity between sperm surface beta-1,4-galactosyltransferase and egg-coat ZP3 mediates sperm-egg binding. Nature 397 1608469
1995 Oocyte Gal alpha 1,3Gal epitopes implicated in sperm adhesion to the zona pellucida glycoprotein ZP3 are not required for fertilization in the mouse. The Journal of biological chemistry 314 7545161
1997 Zp3-cre, a transgenic mouse line for the activation or inactivation of loxP-flanked target genes specifically in the female germ line. Current biology : CB 306 9016703
1989 95 kd sperm proteins bind ZP3 and serve as tyrosine kinase substrates in response to zona binding. Cell 301 2472220
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. Proceedings of the National Academy of Sciences of the United States of America 299 2842789
1992 A large domain common to sperm receptors (Zp2 and Zp3) and TGF-beta type III receptor. FEBS letters 285 1313375
1994 Cloning and characterization of zona pellucida genes and cDNAs from a variety of mammalian species: the ZPA, ZPB and ZPC gene families. DNA sequence : the journal of DNA sequencing and mapping 273 7841460
1996 Mice homozygous for an insertional mutation in the Zp3 gene lack a zona pellucida and are infertile. Development (Cambridge, England) 265 8787763
2001 Trp2 regulates entry of Ca2+ into mouse sperm triggered by egg ZP3. Nature cell biology 254 11331878
1990 Identification of a ZP3-binding protein on acrosome-intact mouse sperm by photoaffinity crosslinking. Proceedings of the National Academy of Sciences of the United States of America 196 2371290
1989 Evidence that aggregation of mouse sperm receptors by ZP3 triggers the acrosome reaction. The Journal of cell biology 185 2544604
2000 Ca(2+) entry through store-operated channels in mouse sperm is initiated by egg ZP3 and drives the acrosome reaction. Molecular biology of the cell 184 10793136
1992 Autoimmune disease of the ovary induced by a ZP3 peptide from the mouse zona pellucida. The Journal of clinical investigation 174 1370297
1988 Molecular analysis of cDNA coding for ZP3, a sperm binding protein of the mouse zona pellucida. Developmental biology 159 3378665
1987 Oocyte-specific expression and developmental regulation of ZP3, the sperm receptor of the mouse zona pellucida. Developmental biology 143 2884155
1994 Sperm-egg recognition in the mouse: characterization of sp56, a sperm protein having specific affinity for ZP3. The Journal of cell biology 140 8188752
1998 Hetero-oligomerization-dependent binding of pig oocyte zona pellucida glycoproteins ZPB and ZPC to boar sperm membrane vesicles. The Journal of biological chemistry 126 9516448
1994 Recombinant human zona pellucida protein ZP3 produced by chinese hamster ovary cells induces the human sperm acrosome reaction and promotes sperm-egg fusion. Biology of reproduction 126 7819440
2010 Insights into egg coat assembly and egg-sperm interaction from the X-ray structure of full-length ZP3. Cell 115 20970175
1996 ZP3-dependent activation of sperm cation channels regulates acrosomal secretion during mammalian fertilization. The Journal of cell biology 109 8707844
2008 Crystal structure of the ZP-N domain of ZP3 reveals the core fold of animal egg coats. Nature 106 19052627
2017 A Recurrent Missense Mutation in ZP3 Causes Empty Follicle Syndrome and Female Infertility. American journal of human genetics 104 28886344
2019 Novel mutations in ZP1, ZP2, and ZP3 cause female infertility due to abnormal zona pellucida formation. Human genetics 97 30810869
1995 Mapping the mouse ZP3 combining site for sperm by exon swapping and site-directed mutagenesis. Proceedings of the National Academy of Sciences of the United States of America 91 7816829
1992 Analysis of the contraceptive potential of antibodies against native and deglycosylated porcine ZP3 in vivo and in vitro. Biology of reproduction 89 1576251
1990 An upstream region of the mouse ZP3 gene directs expression of firefly luciferase specifically to growing oocytes in transgenic mice. Proceedings of the National Academy of Sciences of the United States of America 84 2402504
2010 Genetic evidence for Dnmt3a-dependent imprinting during oocyte growth obtained by conditional knockout with Zp3-Cre and complete exclusion of Dnmt3b by chimera formation. Genes to cells : devoted to molecular & cellular mechanisms 83 20132320
1993 Nucleotide sequence of cDNA encoding ZP3 alpha, a sperm-binding glycoprotein from zona pellucida of pig oocyte. Biochimica et biophysica acta 83 8357839
1996 Molecular cloning, structural analysis, and expression of carp ZP3 gene. Molecular reproduction and development 80 8858599
2017 Dosage effects of ZP2 and ZP3 heterozygous mutations cause human infertility. Human genetics 78 28646452
1999 A 42-kDa glycoprotein from chicken egg-envelope, an avian homolog of the ZPC family glycoproteins in mammalian Zona pellucida. Its first identification, cDNA cloning and granulosa cell-specific expression. European journal of biochemistry 77 10103002
1992 Recombinant mouse ZP3 inhibits sperm binding and induces the acrosome reaction. Developmental biology 76 1577197
1991 Embryonal carcinoma cells transfected with ZP3 genes differentially glycosylate similar polypeptides and secrete active mouse sperm receptor. The Journal of cell biology 76 1655811
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