Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line

Mirjana Martic, Eric K. Moses, Tim E. Adams, De Yi Liu, Debra A. Gook, Claire Garrett, Marjorie E. Dunlop, Gordon H.W. Baker

    Research output: Contribution to journalArticle

    14 Scopus citations

    Abstract

    Aim: To produce biologically active recombinant human (rh) ZP proteins in a human cell for use in sperm function tests. Methods: The human embryonic kidney cell line 293T was employed to produce rhZP1, rhZP2 and rhZP3 proteins individually and together by co-expression. Presence of these proteins in the culture medium and cell lysate was assessed by Western blotting analysis. The effect of the recombinant proteins on the human AR was assessed. Results: RhZP2 and rhZP3 were secreted into the culture medium, whereas rhZP1 was found only in the cell lysate. Interestingly, when all zona pellucida proteins were co-expressed in the same cells, rhZP1 was also secreted into the culture medium. However, despite the presence of all three ZP proteins in sufficient concentration and evidence of heavy glycosylation on gel electrophoresis, biological activity to induce the AR was not observed. Conclusion: RhZP1, rhZP2 and rhZP3 were successfully expressed in the human embryonic kidney cell line 293T. It appears that an interaction amongst these proteins may be required for release of rhZP1 from the cell. Although this approach is not satisfactory for producing active human ZP proteins, it makes a significant contribution to the understanding of the structural and functional characteristics of the ZP proteins.

    Original languageEnglish (US)
    Pages (from-to)3-13
    Number of pages11
    JournalAsian Journal of Andrology
    Volume6
    Issue number1
    StatePublished - Mar 1 2004

    Keywords

    • Acrosome reaction
    • Glycosylation
    • Human cell line
    • Recombinant proteins
    • Zona pellucida

    ASJC Scopus subject areas

    • Urology

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  • Cite this

    Martic, M., Moses, E. K., Adams, T. E., Liu, D. Y., Gook, D. A., Garrett, C., Dunlop, M. E., & Baker, G. H. W. (2004). Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line. Asian Journal of Andrology, 6(1), 3-13.