Letter | Published:

Crystal structure of the ZP-N domain of ZP3 reveals the core fold of animal egg coats

Nature volume 456, pages 653657 (04 December 2008) | Download Citation

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Abstract

Species-specific recognition between the egg extracellular matrix (zona pellucida) and sperm is the first, crucial step of mammalian fertilization1. Zona pellucida filament components ZP3 and ZP2 act as sperm receptors, and mice lacking either of the corresponding genes produce oocytes without a zona pellucida and are completely infertile2. Like their counterparts in the vitelline envelope of non-mammalian eggs and many other secreted eukaryotic proteins, zona pellucida subunits polymerize using a ‘zona pellucida (ZP) domain’ module3,4,5, whose conserved amino-terminal part (ZP-N) was suggested to constitute a domain of its own6. No atomic structure has been reported for ZP domain proteins, and there is no structural information on any conserved vertebrate protein that is essential for fertilization and directly involved in egg–sperm binding. Here we describe the 2.3 ångström (Å) resolution structure of the ZP-N fragment of mouse primary sperm receptor ZP3. The ZP-N fold defines a new immunoglobulin superfamily subtype with a β-sheet extension characterized by an E′ strand and an invariant tyrosine residue implicated in polymerization. The structure strongly supports the presence of ZP-N repeats within the N-terminal region of ZP2 and other vertebrate zona pellucida/vitelline envelope proteins, with implications for overall egg coat architecture, the post-fertilization block to polyspermy and speciation. Moreover, it provides an important framework for understanding human diseases caused by mutations in ZP domain proteins and developing new methods of non-hormonal contraception.

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Accessions

Primary accessions

Data deposits

Atomic coordinates and structure factors have been deposited with the Protein Data Bank under accession codes 3D4C (crystal form I), 3D4G (crystal form II) and 3EF7 (crystal form III).

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Acknowledgements

Acknowledgments This work was supported by Karolinska Institutet, the Swedish Research Council (grant 2005-5102) and European Community (Marie Curie ERG 31055). We thank P. Nordlund and G. Schneider for access to the Stockholm SGC robotic crystallization facility and beamtime; H. Belrhali, J. McCarthy, R. Ravelli and M. Walsh for assistance at ESRF; P. Afonine, R. Grosse-Kunstleve and P. Zwart for help with phenix.refine; G. Murshudov for help with REFMAC; C. Chothia, F. Cotelli, J.-Å. Gustafsson, R. Herbst-Irmer, A. Kohl, R. Ladenstein, M. Letarte, E. Litscher, E. Morgunova, A. Murzin, K. Nagai, L. Nilsson, D. Rhodes, R. Toftgård and P. Wassarman for discussions and comments.

Author Contributions M.M., L.H. and L.J. generated constructs and performed protein expression and purification. T.S. performed mass spectrometric analysis. S.B. carried out molecular dynamics simulations. L.J. and M.M. crystallized proteins, determined structures and wrote the manuscript.

Author information

Author notes

    • Magnus Monné
    •  & Ling Han

    These authors contributed equally to this work.

Affiliations

  1. Karolinska Institutet, Department of Biosciences and Nutrition, Hälsovägen 7, SE-141 57 Huddinge, Sweden

    • Magnus Monné
    • , Ling Han
    • , Thomas Schwend
    • , Sofia Burendahl
    •  & Luca Jovine

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Correspondence to Luca Jovine.

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https://doi.org/10.1038/nature07599

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