Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:Q07644 (polypeptide)
 72,197 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The distribution of O-linked oligosaccharides on the M(r) 55,000 glycoproteins, ZP3 alpha and ZP3 beta, of the porcine oocyte zona pellucida was examined. Purified preparations of endo-beta-galactosidase digested ZP3 alpha and ZP3 beta were reduced and carboxamidomethylated and digested with trypsin. When the trypsin digests were mapped by HPLC, each glycoprotein yielded only one N-acetylgalactosamine containing glycopeptide. Purification of the O-glycopeptides was achieved by a two-step protocol. Tryptic digests were applied to jacalin-agarose and specifically-bound O-glycopeptides (alpha OGP and beta OGP) were eluted with buffer containing 50 mM alpha-methylgalactoside as the haptenic sugar. Further purification of each O-glycopeptide was accomplished by reverse phase HPLC. Purified O-glycopeptides were characterized with respect to amino acid and carbohydrate compositions and sequenced by automated Edman degradation; alpha OGP was a 41-residue glycopeptide with three O-linked sugar chains. Sequence comparisons revealed a 75% identity between alpha OGP and a corresponding segment of rabbit rec55 zona protein; beta OGP was a 25-residue glycopeptide characterized by the presence of one N-linked and five O-linked sugar chains and a trypsin-resistant internal arginine residue. Sequence alignments revealed an 80% or greater identity between beta OGP and internal peptides of mouse, hamster and human ZP3 zona proteins. These studies demonstrate that in the case of ZP3 alpha and ZP3 beta, the pig homologues of rabbit rec55 and mouse ZP3, respectively, O-linked oligosaccharides are confined within delimited domains rather than widely dispersed on the polypeptide backbone. Such clustering of O-linked oligosaccharides may represent an essential determinant of the structure and biological activity of zona proteins.
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PMID:Porcine oocyte zona pellucida M(r) 55,000 glycoproteins: identification of O-glycosylated domains. 141 87

We have undertaken a comparative analysis of the contraceptive activity of antibodies directed against the porcine sperm receptor zona pellucida antigen (ZP3) and its Mr = 32,000 polypeptide core (DGZP-32). The strategies employed for this analysis included the induction of active immunity in a primate, the common marmoset, and an in vitro fertilization protocol involving the use of viable human ova. In both experimental situations, antibodies against ZP3 were shown to exhibit contraceptive activity, leading respectively to the induction of long-term infertility in the primate model and to the complete inhibition of human fertilization in vitro. The in vivo studies also revealed that the induction of high titer antibodies against ZP3 was inevitably associated with the appearance of an ovarian pathology characterized by the progressive depletion of the primordial follicle pool within one to two years. This side effect could not be alleviated by the use of DGZP-32 as antigen since the induction of immunity against this polypeptide was also associated with the eventual appearance of an ovarian pathology identical to that observed with ZP3. Furthermore, the DGZP-32 peptide was less effective than ZP3 in inducing the formation of antibodies capable of inhibiting the fertilization of human ova in vitro. We conclude that significant problems remain with the use of deglycosylated zona peptides for the development of contraceptive vaccines and that their potential will not be realized until the epitopes responsible for the induction of infertility and the primordial follicle depletion have been identified and segregated.
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PMID:Analysis of the contraceptive potential of antibodies against native and deglycosylated porcine ZP3 in vivo and in vitro. 157 51

Hamster zonae pellucidae were obtained from follicular oocytes, superovulated eggs, and eggs fertilized in vivo or in vitro. Zonae were labelled with N-succinimidyl-3(4-hydroxy,5-[125I]iodophenyl)propionate, and compared on single- and two-dimensional SDS-PAGE. Single-dimensional electrophoresis showed considerable differences between zona categories in the amount of label that they incorporated; follicular zonae incorporated the least label and zonae from eggs fertilized in vivo the most. On two-dimensional electrophoresis, polypeptides from 3 of the 4 zona categories migrated into 4 major groups: two of these groups each with Mr 150,000-250,000 were within the Mr range of ZP1, and two others, at Mr 90,000 and 55,000, appeared to be analogous to ZP2 and ZP3, respectively. The fourth zona category (zonae from eggs fertilized in vivo) showed a changed polypeptide profile as well as incorporating the most label; one of the polypeptides, Mr 150,000-250,000, was undetectable, but a train of Mr 70,000-90,000 polypeptides and a discrete polypeptide at Mr 20,000 were new. Since this changed profile did not occur in zonae from superovulated eggs, or in zonae from eggs fertilized in vitro, a synergism between oviductal factors and factors from the spermatozoon or egg, or both, towards the zona in vivo is indicated.
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PMID:Changes in the composition of the hamster zona pellucida after fertilization in vivo but not in vitro. 225 Feb 44

During the course of fertilization in mammals, free-swimming sperm bind tightly to receptors located in the egg extracellular coat, or zona pellucida. Recently, the hamster sperm receptor, a 56,000 Mr zona pellucida glycoprotein called hZP3, was identified and partially characterized (C. C. Moller et al., (1990). Dev. Biol. 137, 276-286). Here, we describe genomic cloning of hZP3, certain organizational features of the hZP3 gene, and primary structures of hZP3 mRNA and polypeptide. The findings are compared with reported results of comparable analyses of the mouse sperm receptor, an 83,000 Mr zona pellucida glycoprotein called mZP3. Such comparisons reveal a high degree of conservation of genomic organization and polypeptide structure for the two mammalian sperm receptors, despite the considerable difference in their Mrs. These findings are of interest in view of the extremely restricted expression of the ZP3 gene during development and the important role of ZP3 oligosaccharides in gamete adhesion.
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PMID:Genomic organization and polypeptide primary structure of zona pellucida glycoprotein hZP3, the hamster sperm receptor. 185 Mar 70

The mouse sperm receptor, called ZP3, is a glycoprotein (83,000 Mr) that consists of a 44,000 Mr polypeptide chain (402 amino acids), three or four N-linked oligosaccharides, and an undetermined number of O-linked oligosaccharides. There are more than 10(9) copies of ZP3 present throughout the mouse egg extracellular coat, or zona pellucida. As a prelude to fertilization, each acrosome-intact sperm binds in a relatively species-specific manner to tens-of-thousands of copies of ZP3 at the surface of the zona pellucida. Binding to ZP3 induces sperm to undergo the acrosome reaction (membrane fusion) and, consequently, enables them to penetrate through the zona pellucida and to reach, and then fuse with, egg plasma membrane (fertilization). Purified ZP3, as well as a specific class of ZP3-derived O-linked oligosaccharides (3900 Mr), exhibit sperm receptor activity in vitro. The oligosaccharides, which represent a relatively low percentage of total ZP3 O-linked oligosaccharides, account for the glycoprotein's sperm receptor activity in vitro (i.e., recognition and binding). Furthermore, either enzymic removal or modification of certain sugars that constitute these oligosaccharides results in destruction of sperm receptor activity. These and other findings strongly suggest that during mammalian fertilization carbohydrates play a fundamental role in species-specific sperm-egg interactions.
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PMID:Role of carbohydrates in receptor-mediated fertilization in mammals. 255 6

ZP3, a glycoprotein present in the extracellular coat (zona pellucida) of the unfertilized egg, serves as a receptor for sperm and an inducer of the acrosome reaction (sperm exocytosis) during fertilization in mice. As such, ZP3 regulates the initial species-specific interactions between male and female mouse gametes. Recently, the organization and expression of the gene encoding ZP3 have been studied in some detail. These studies have led to some important findings, including the entire primary structure of the glycoprotein's polypeptide chain and the sequence of more than 11 kilobases of DNA at the ZP3 genomic locus. The latter includes the entire transcription unit for ZP3, as well as 5' and 3' flanking sequences. Of particular interest is the finding that the ZP3 gene is expressed at extremely high levels by growing oocytes and by no other cell type in the mouse. This oocyte-specific expression occurs only at a particular stage of oogenesis. Although the specific regulatory elements responsible for the highly restricted expression of ZP3 have not been identified as yet, certain organizational features of the ZP3 gene that have been described may be relevant in this connection. Further molecular analyses of ZP3 will provide additional insight into its synthesis, structure, and functions, and could have practical consequences in the context of human conception and contraception.
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PMID:Profile of a mammalian sperm receptor gene. 256 22

The initial communication between the gametes is a molecular, receptor-mediated process that takes place at the surface of the egg coat. The zona pellucida plays a central role in this process such that, on the one hand, spermatozoa may bind to it and, on the other hand, it prevents polyspermy. In the mouse, ZP3, a glycoprotein of the zona pellucida with a mol. wt of 84 kd, serves as a sperm receptor. Only a relatively small part of ZP3, namely certain O-linked carbohydrate side chains, is involved in the process of binding. These oligosaccharides probably become bound to enzymes associated with the plasma membrane of the sperm head and thus form an enzyme-substrate complex. A terminal alpha-galactose has been found to be one of the decisive sugar molecules and, moreover, the critical chemical group. After sperm binding to the zona pellucida has taken place, the polypeptide chain of ZP3 initiates the acrosome reaction in the sperm head. In the mouse, numerous binding proteins have been detected in the sperm plasma membrane: these are enzymes such as glycosyl transferases, proteinases, and glycosidases. A galactosyl transferase has been found on the surface of the mouse sperm that binds specifically to N-acetylglucosamine in the mouse zona pellucida. It is therefore apparent that carbohydrate-binding proteins on the sperm surface mediate gamete recognition through their high affinity and specificity for complex glycoconjugates in the egg coat. In fact, it is not at all surprising that complementary cell-surface protein and glycoconjugates are involved in fertilization, since many somatic cells exhibit a similar mechanism of cell recognition.
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PMID:Molecular aspects of mammalian fertilization. 269 82

The mouse sperm receptor, a glycoprotein called ZP3, is synthesized and secreted by growing oocytes. It is present in more than a billion copies in the unfertilized egg's extracellular coat, or zona pellucida. We have cloned and characterized a region of the mouse (CD-1) genome that spans 10 kilobases of the ZP3 locus. The genomic clones described encompass the entire ZP3 coding region, which contains eight exons. The exons were identified, mapped, and sequenced, yielding the entire primary structure of the ZP3 polypeptide chain (424 amino acids; Mr, 46,300), which includes a 22-amino acid signal sequence. In addition, sequencing of genomic clones has revealed some unusual features of ZP3 mRNA and a region just downstream of the ZP3 gene.
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PMID:Primary structure of the mouse sperm receptor polypeptide determined by genomic cloning. 284 70

Previously, we reported that ZP3, one of three different glycoproteins present in the mouse egg's zona pellucida, serves as a sperm receptor. Furthermore, small glycopeptides derived from egg ZP3 retain full sperm receptor activity, suggesting a role for carbohydrate, rather than polypeptide chain in receptor function. Here, we report that removal of O-linked oligosaccharides from ZP3 destroys its sperm receptor activity, whereas removal of N-linked oligosaccharides has no effect. A specific size class of O-linked oligosaccharides, recovered following mild alkaline hydrolysis and reduction of ZP3, is shown to possess sperm receptor activity and to bind to sperm. The results presented strongly suggest that mouse sperm bind to eggs via O-linked oligosaccharides present on ZP3.
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PMID:O-linked oligosaccharides of mouse egg ZP3 account for its sperm receptor activity. 298 49

The major macromolecular component of the porcine oocyte zona pellucida is a Mr = 55,000 antigen, termed ZP3, comprised of greater than 25 charge isomers. ZP3 was purified to apparent electrophoretic homogeneity from nonreduced, sodium dodecyl sulfate-treated porcine zonae pellucidae by chromatography on Sephacryl S-400 and hydroxylapatite resins. The carbohydrate moiety of purified ZP3 was comprised of a heterogeneous population of acidic lactosaminoglycans as evidenced by the saccharide composition and size distribution of glycopeptides produced by Pronase digestion of ZP3, as well as by the sensitivity of ZP3 to digestion with Escherichia freundii endo-beta-galactosidase. Endo-beta-galactosidase-digested ZP3 was resolved by gel electrophoresis into two components, termed alpha-glycoprotein and beta-glycoprotein, with Mr values (nonreduced) of 46,000 and 42,000, respectively. Each was comprised of fewer and more neutral charge isomers than ZP3. Following purification by reverse phase high performance liquid chromatography, the alpha- and beta-glycoproteins of endo-beta-galactosidase-digested ZP3 were distinguished on the basis of amino acid and carbohydrate compositions, amino-terminal sequence analyses and peptide mapping experiments, thus suggesting differences in the primary structures of their respective polypeptide moieties. Corresponding dissimilarities in the immunoreactivities of the alpha- and beta-glycoproteins toward polyclonal antisera raised against ZP3, alpha-glycoprotein, and beta-glycoprotein were revealed by competitive binding radioimmunoassays as well as by immunoblotting experiments. Collectively, the data were interpreted to indicate that the Mr = 55,000 antigen of porcine oocyte zona pellucida is in fact comprised of overlapping families of charge isomers corresponding to two structurally and immunologically distinct lactosaminoglycan-containing glycoproteins.
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PMID:Structural characterization of the Mr = 55,000 antigen (ZP3) of porcine oocyte zona pellucida. Purification and characterization of alpha- and beta-glycoproteins following digestion of lactosaminoglycan with endo-beta-galactosidase. 310 May 14


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