HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Thaler, C. D. Articles by Cardullo, R. A. Search for Related Content PubMed PubMed Citation Articles by Thaler, C. D. Articles by Cardullo, R. A. Social Bookmarking                      What's this?

Volume 271, Number 38, Issue of September 20, 1996 pp. 23289-23297
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

The Initial Molecular Interaction between Mouse Sperm and the Zona Pellucida Is a Complex Binding Event

(Received for publication, March 14, 1996, and in revised form, June 4, 1996)

From the University of California, Department of Biology, Riverside, California 92521

Prior to fertilization, mammalian sperm must first bind to the zona pellucida (ZP), a glycoprotein matrix surrounding the egg. Sperm specifically bind to ZP3, an 83-kDa glycoprotein which functions as both an adhesion molecule and as a secretagogue for acrosomal exocytosis (Litscher, E. S., and Wassarman, P. M. (1993) Trends Glycosci. Glycotechnol. 5, 369-388). We used acid solubilized, ¹²⁵I-labeled ZPs to quantify the initial binding event on mouse spermatozoa. Live sperm could not be used since solubilized ZPs rapidly initiated exocytosis. Instead, acrosome intact mouse sperm were briefly fixed in 1% glutaraldehyde for binding studies using a standard filtration assay. The fixed sperm are suitable for sperm-zona binding assays based on two experiments: 1) incubating either live or fixed sperm in low concentrations of ¹²⁵I-ZPs not sufficient to induce acrosomal exocytosis revealed no differences in binding up to 15 min and 2) solubilized, unlabeled ZPs competed for ¹²⁵I-ZPs with an K_I of approximately 3.78 n. Sperm-¹²⁵I-ZP binding reached equilibrium with a 1/2 of ~22 min at 37 °C. Affinity parameters were calculated using the well substantiated assumption that only ZP3 binds intact mouse sperm. The on-rate constant for association of ¹²⁵I-ZP binding to the mouse sperm surface was calculated to be 3.2 × 10^{6 1} min¹. The saturation binding isotherm revealed that there are approximately 30,000 binding sites, ascribed to ZP3, with an EC₅₀ of 1.29 n. Further analysis indicated that this binding is complex (Hill coefficient = 1.72), suggesting involvement of multiple receptors on the sperm surface and/or multiple ligand moieties. High and low affinity ZP binding sites on the sperm surface were confirmed by dissociation experiments. ¹²⁵I-ZP dissociation was clearly biphasic, and kinetic off-rate constants of 0.161 min¹ and 0.0023 min¹ were calculated for the low and high affinity sites, respectively. Apparent affinities (K_d values) of 50 n for the low affinity and 0.72 n for the high affinity interaction were calculated from the rate constants. These data demonstrate that the initial adhesion event between mouse sperm and the zona pellucida is a high affinity event which is sufficient to tether a sperm to the extracellular matrix prior to the induction of acrosomal exocytosis.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				D.Y. Liu, M.L. Liu, and H.W.G. Baker Enhancement of sperm-zona pellucida (ZP) binding capacity by activation of protein kinase A and C pathways in certain infertile men with defective sperm-ZP binding Hum. Reprod., January 1, 2009; 24(1): 20 - 27. [Abstract] [Full Text] [PDF]

				P. C.N. Chiu, B. S.T. Wong, C.L. Lee, R. T.K. Pang, K.-F. Lee, S.B. Sumitro, S.K. Gupta, and W. S.B. Yeung Native human zona pellucida glycoproteins: purification and binding properties Hum. Reprod., June 1, 2008; 23(6): 1385 - 1393. [Abstract] [Full Text] [PDF]

				A. Walsh, D. Whelan, A. Bielanowicz, B. Skinner, R. J. Aitken, M. K. O'Bryan, and B. Nixon Identification of the Molecular Chaperone, Heat Shock Protein 1 (Chaperonin 10), in the Reproductive Tract and in Capacitating Spermatozoa in the Male Mouse Biol Reprod, June 1, 2008; 78(6): 983 - 993. [Abstract] [Full Text] [PDF]

				J. A. Riffell and R. K. Zimmer Sex and flow: the consequences of fluid shear for sperm egg interactions J. Exp. Biol., October 15, 2007; 210(20): 3644 - 3660. [Abstract] [Full Text] [PDF]

				R. A. van Gestel, I. A. Brewis, P. R. Ashton, J. F. Brouwers, and B. M. Gadella Multiple proteins present in purified porcine sperm apical plasma membranes interact with the zona pellucida of the oocyte Mol. Hum. Reprod., July 1, 2007; 13(7): 445 - 454. [Abstract] [Full Text] [PDF]

				P. C. N. Chiu, M.-K. Chung, R. Koistinen, H. Koistinen, M. Seppala, P.-C. Ho, E. H. Y. Ng, K.-F. Lee, and W. S. B. Yeung Glycodelin-A interacts with fucosyltransferase on human sperm plasma membrane to inhibit spermatozoa-zona pellucida binding J. Cell Sci., January 1, 2007; 120(1): 33 - 44. [Abstract] [Full Text] [PDF]

				R.A. van Gestel, I.A. Brewis, P.R. Ashton, J.B. Helms, J.F. Brouwers, and B.M. Gadella Capacitation-dependent concentration of lipid rafts in the apical ridge head area of porcine sperm cells Mol. Hum. Reprod., August 1, 2005; 11(8): 583 - 590. [Abstract] [Full Text] [PDF]

				N. Yonezawa, K. Kudo, H. Terauchi, S. Kanai, N. Yoda, M. Tanokura, K. Ito, K.-i. Miura, T. Katsumata, and M. Nakano Recombinant Porcine Zona Pellucida Glycoproteins Expressed in Sf9 Cells Bind to Bovine Sperm but Not to Porcine Sperm J. Biol. Chem., May 27, 2005; 280(21): 20189 - 20196. [Abstract] [Full Text] [PDF]

				K. L. Asquith, A. J. Harman, E. A. McLaughlin, B. Nixon, and R. J. Aitken Localization and Significance of Molecular Chaperones, Heat Shock Protein 1, and Tumor Rejection Antigen gp96 in the Male Reproductive Tract and During Capacitation and Acrosome Reaction Biol Reprod, February 1, 2005; 72(2): 328 - 337. [Abstract] [Full Text] [PDF]

				P.C.N. Chiu, H.Y. Tsang, R. Koistinen, H. Koistinen, M. Seppala, K.F. Lee, and W.S.B. Yeung The Contribution of D-Mannose, L-Fucose, N-Acetylglucosamine, and Selectin Residues on the Binding of Glycodelin Isoforms to Human Spermatozoa Biol Reprod, June 1, 2004; 70(6): 1710 - 1719. [Abstract] [Full Text] [PDF]

				T. L. Tollner, A. I. Yudin, G. N. Cherr, and J. W. Overstreet Real-Time Observations of Individual Macaque Sperm Undergoing Tight Binding and the Acrosome Reaction on the Zona Pellucida Biol Reprod, February 1, 2003; 68(2): 664 - 672. [Abstract] [Full Text] [PDF]

				C. L. Kerr, W. F. Hanna, J. H. Shaper, and W. W. Wright Characterization of Zona Pellucida Glycoprotein 3 (ZP3) and ZP2 Binding Sites on Acrosome-Intact Mouse Sperm Biol Reprod, June 1, 2002; 66(6): 1585 - 1595. [Abstract] [Full Text] [PDF]

				D. J. Miller, X. Shi, and H. Burkin Molecular Basis of Mammalian Gamete Binding Recent Prog. Horm. Res., January 1, 2002; 57(1): 37 - 73. [Abstract] [Full Text] [PDF]

				S. S. Baker, R. A. Cardullo, and C. D. Thaler Sonication of Mouse Sperm Membranes Reveals Distinct Protein Domains Biol Reprod, January 1, 2002; 66(1): 57 - 64. [Abstract] [Full Text]

				C. D. Thaler and R. A. Cardullo Distinct Membrane Fractions from Mouse Sperm Bind Different Zona Pellucida Glycoproteins Biol Reprod, January 1, 2002; 66(1): 65 - 69. [Abstract] [Full Text]

				K. J. Mengerink and V. D. Vacquier Glycobiology of sperm-egg interactions in deuterostomes Glycobiology, April 1, 2001; 11(4): 37R - 43R. [Abstract] [Full Text] [PDF]

				C. R. Loeser, C. I. Lynch, and D. R.P. Tulsiani Characterization of the Pharmacological-Sensitivity Profile of Neoglycoprotein-Induced Acrosome Reaction in Mouse Spermatozoa Biol Reprod, September 1, 1999; 61(3): 629 - 634. [Abstract] [Full Text]

				C. R. Loeser and D. R.P. Tulsiani The Role of Carbohydrates in the Induction of the Acrosome Reaction in Mouse Spermatozoa Biol Reprod, January 1, 1999; 60(1): 94 - 101. [Abstract] [Full Text]

				L.-Y. Chen, Y.-H. Lin, M.-L. Lai, and Y.-H. Chen Developmental Profile of a Caltrin-Like Protease Inhibitor, P12, in Mouse Seminal Vesicle and Characterization of Its Binding Sites on Sperm Surface Biol Reprod, December 1, 1998; 59(6): 1498 - 1505. [Abstract] [Full Text]

				E. C. Yurewicz, A. G. Sacco, S. K. Gupta, N. Xu, and D. A. Gage Hetero-oligomerization-dependent Binding of Pig Oocyte Zona Pellucida Glycoproteins ZPB and ZPC to Boar Sperm Membrane Vesicles J. Biol. Chem., March 27, 1998; 273(13): 7488 - 7494. [Abstract] [Full Text] [PDF]

				D. S. Johnston, W. W. Wright, J. H. Shaper, C. H. Hokke, D. H. Van den Eijnden, and D. H. Joziasse Murine Sperm-Zona Binding, A Fucosyl Residue Is Required for a High Affinity Sperm-binding Ligand. A SECOND SITE ON SPERM BINDS A NONFUCOSYLATED, beta -GALACTOSYL-CAPPED OLIGOSACCHARIDE J. Biol. Chem., January 23, 1998; 273(4): 1888 - 1895. [Abstract] [Full Text] [PDF]

				W. J. Swanson and V. D. Vacquier The abalone egg vitelline envelope receptor for sperm lysin is a giant multivalent molecule PNAS, June 24, 1997; 94(13): 6724 - 6729. [Abstract] [Full Text] [PDF]

				C.-W. Luo, H.-J. Lin, and Y.-H. Chen A Novel Heat-labile Phospholipid-binding Protein, SVS VII, in Mouse Seminal Vesicle as a Sperm Motility Enhancer J. Biol. Chem., March 2, 2001; 276(10): 6913 - 6921. [Abstract] [Full Text] [PDF]