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. 1990 Jul;87(14):5563–5567. doi: 10.1073/pnas.87.14.5563

Identification of a ZP3-binding protein on acrosome-intact mouse sperm by photoaffinity crosslinking.

J D Bleil 1, P M Wassarman 1
PMCID: PMC54365  PMID: 2371290

Abstract

During the process of fertilization in mammals, sperm bind in a relatively species-specific manner to the zona pellucida (ZP) of ovulated eggs. ZP3, a glycoprotein found in the mouse egg zona pellucida, serves as receptor for sperm during gamete adhesion. We report here that a Mr 56,000 protein found on mouse sperm has properties expected for a sperm component that recognizes and binds to ZP3. This sperm protein is radiolabeled preferentially by a photoactivatable heterobifunctional crosslinker ("Denny-Jaffee reagent") covalently linked to purified ZP3, binds very tightly to ZP3-affinity columns, and is localized to heads of acrosome-intact but not acrosome-reacted sperm. These and other findings suggest that this protein may be a "ZP3-binding protein" that, together with the sperm receptor, supports species-specific binding of mouse sperm to unfertilized eggs.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abdullah M., Kierszenbaum A. L. Identification of rat testis galactosyl receptor using antibodies to liver asialoglycoprotein receptor: purification and localization on surfaces of spermatogenic cells and sperm. J Cell Biol. 1989 Feb;108(2):367–375. doi: 10.1083/jcb.108.2.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aguas A. P., da Silva P. P. Bimodal redistribution of surface transmembrane glycoproteins during Ca2+-dependent secretion (acrosome reaction) in boar spermatozoa. J Cell Sci. 1989 Jul;93(Pt 3):467–479. doi: 10.1242/jcs.93.3.467. [DOI] [PubMed] [Google Scholar]
  3. Benau D. A., Storey B. T. Characterization of the mouse sperm plasma membrane zona-binding site sensitive to trypsin inhibitors. Biol Reprod. 1987 Mar;36(2):282–292. doi: 10.1095/biolreprod36.2.282. [DOI] [PubMed] [Google Scholar]
  4. Bleil J. D., Greve J. M., Wassarman P. M. Identification of a secondary sperm receptor in the mouse egg zona pellucida: role in maintenance of binding of acrosome-reacted sperm to eggs. Dev Biol. 1988 Aug;128(2):376–385. doi: 10.1016/0012-1606(88)90299-0. [DOI] [PubMed] [Google Scholar]
  5. Bleil J. D., Wassarman P. M. Autoradiographic visualization of the mouse egg's sperm receptor bound to sperm. J Cell Biol. 1986 Apr;102(4):1363–1371. doi: 10.1083/jcb.102.4.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bleil J. D., Wassarman P. M. 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 U S A. 1988 Sep;85(18):6778–6782. doi: 10.1073/pnas.85.18.6778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bleil J. D., Wassarman P. M. Mammalian sperm-egg interaction: identification of a glycoprotein in mouse egg zonae pellucidae possessing receptor activity for sperm. Cell. 1980 Jul;20(3):873–882. doi: 10.1016/0092-8674(80)90334-7. [DOI] [PubMed] [Google Scholar]
  8. Brown C. R., Jones R. Binding of zona pellucida proteins to a boar sperm polypeptide of Mr 53,000 and identification of zona moieties involved. Development. 1987 Mar;99(3):333–339. doi: 10.1242/dev.99.3.333. [DOI] [PubMed] [Google Scholar]
  9. Denny J. B., Blobel G. 125I-labeled crosslinking reagent that is hydrophilic, photoactivatable, and cleavable through an azo linkage. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5286–5290. doi: 10.1073/pnas.81.17.5286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Florman H. M., Bechtol K. B., Wassarman P. M. Enzymatic dissection of the functions of the mouse egg's receptor for sperm. Dev Biol. 1984 Nov;106(1):243–255. doi: 10.1016/0012-1606(84)90079-4. [DOI] [PubMed] [Google Scholar]
  11. Florman H. M., Storey B. T. Mouse gamete interactions: the zona pellucida is the site of the acrosome reaction leading to fertilization in vitro. Dev Biol. 1982 May;91(1):121–130. doi: 10.1016/0012-1606(82)90015-x. [DOI] [PubMed] [Google Scholar]
  12. Florman H. M., Wassarman P. M. O-linked oligosaccharides of mouse egg ZP3 account for its sperm receptor activity. Cell. 1985 May;41(1):313–324. doi: 10.1016/0092-8674(85)90084-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gao B., Klein L. E., Britten R. J., Davidson E. H. Sequence of mRNA coding for bindin, a species-specific sea urchin sperm protein required for fertilization. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8634–8638. doi: 10.1073/pnas.83.22.8634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Glabe C. G., Grabel L. B., Vacquier V. D., Rosen S. D. Carbohydrate specificity of sea urchin sperm bindin: a cell surface lectin mediating sperm-egg adhesion. J Cell Biol. 1982 Jul;94(1):123–128. doi: 10.1083/jcb.94.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jaffe C. L., Lis H., Sharon N. New clevable photoreactive heterobifunctional cross-linking reagents for studying membrane organization. Biochemistry. 1980 Sep 16;19(19):4423–4429. doi: 10.1021/bi00560a007. [DOI] [PubMed] [Google Scholar]
  16. Leyton L., Saling P. 95 kd sperm proteins bind ZP3 and serve as tyrosine kinase substrates in response to zona binding. Cell. 1989 Jun 30;57(7):1123–1130. doi: 10.1016/0092-8674(89)90049-4. [DOI] [PubMed] [Google Scholar]
  17. Leyton L., Saling P. Evidence that aggregation of mouse sperm receptors by ZP3 triggers the acrosome reaction. J Cell Biol. 1989 Jun;108(6):2163–2168. doi: 10.1083/jcb.108.6.2163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Macek M. B., Shur B. D. Protein-carbohydrate complementarity in mammalian gamete recognition. Gamete Res. 1988 May;20(1):93–109. doi: 10.1002/mrd.1120200109. [DOI] [PubMed] [Google Scholar]
  19. Moller C. C., Bleil J. D., Kinloch R. A., Wassarman P. M. Structural and functional relationships between mouse and hamster zona pellucida glycoproteins. Dev Biol. 1990 Feb;137(2):276–286. doi: 10.1016/0012-1606(90)90254-g. [DOI] [PubMed] [Google Scholar]
  20. O'Rand M. G., Matthews J. E., Welch J. E., Fisher S. J. Identification of zona binding proteins of rabbit, pig, human, and mouse spermatozoa on nitrocellulose blots. J Exp Zool. 1985 Sep;235(3):423–428. doi: 10.1002/jez.1402350314. [DOI] [PubMed] [Google Scholar]
  21. Primakoff P., Hyatt H., Myles D. G. A role for the migrating sperm surface antigen PH-20 in guinea pig sperm binding to the egg zona pellucida. J Cell Biol. 1985 Dec;101(6):2239–2244. doi: 10.1083/jcb.101.6.2239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ram P. A., Cardullo R. A., Millette C. F. Expression and topographical localization of cell surface fucosyltransferase activity during epididymal sperm maturation in the mouse. Gamete Res. 1989 Mar;22(3):321–332. doi: 10.1002/mrd.1120220309. [DOI] [PubMed] [Google Scholar]
  23. Saling P. M. Involvement of trypsin-like activity in binding of mouse spermatozoa to zonae pellucidae. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6231–6235. doi: 10.1073/pnas.78.10.6231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tulsiani D. R., Skudlarek M. D., Orgebin-Crist M. C. Novel alpha-D-mannosidase of rat sperm plasma membranes: characterization and potential role in sperm-egg interactions. J Cell Biol. 1989 Sep;109(3):1257–1267. doi: 10.1083/jcb.109.3.1257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Töpfer-Petersen E., Friess A. E., Nguyen H., Schill W. B. Evidence for a fucose-binding protein in boar spermatozoa. Histochemistry. 1985;83(2):139–145. doi: 10.1007/BF00495144. [DOI] [PubMed] [Google Scholar]
  26. Vacquier V. D., Moy G. W. Isolation of bindin: the protein responsible for adhesion of sperm to sea urchin eggs. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2456–2460. doi: 10.1073/pnas.74.6.2456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wassarman P. M., Bleil J. D., Florman H. M., Greve J. M., Roller R. J., Salzmann G. S., Samuels F. G. The mouse egg's receptor for sperm: what is it and how does it work? Cold Spring Harb Symp Quant Biol. 1985;50:11–19. doi: 10.1101/sqb.1985.050.01.004. [DOI] [PubMed] [Google Scholar]
  28. Wassarman P. M. Early events in mammalian fertilization. Annu Rev Cell Biol. 1987;3:109–142. doi: 10.1146/annurev.cb.03.110187.000545. [DOI] [PubMed] [Google Scholar]
  29. Wassarman P. M. Profile of a mammalian sperm receptor. Development. 1990 Jan;108(1):1–17. doi: 10.1242/dev.108.Supplement.1. [DOI] [PubMed] [Google Scholar]
  30. Wassarman P. M. The biology and chemistry of fertilization. Science. 1987 Jan 30;235(4788):553–560. doi: 10.1126/science.3027891. [DOI] [PubMed] [Google Scholar]

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