Native human zona pellucida glycoproteins: purification and binding properties

Authors: Chiu, Philip C.N.; Wong, Ben S.T.; Lee, C.L.; Pang, Ronald T.K.; Lee, Kai-Fai; Sumitro, S.B.; Gupta, S.K.; Yeung, William S.B.

Source: Human Reproduction, Volume 23, Number 6, 21 June 2008 , pp. 1385-1393(9)

Publisher: Oxford University Press

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Abstract:

BACKGROUND

Fertilization starts with the binding of the spermatozoa to the zona pellucida (ZP) of the oocyte. Such binding is a carbohydrate-mediated event and consists of a series of tightly regulated events. Molecular interactions between spermatozoon and ZP in human are not well characterized due to limited availability of oocytes for research. Our current technology cannot generate recombinant human ZP (hZP) glycoproteins with native glycosylation.

METHODS AND RESULTS

In this study, hZP glycoproteins, hZP2 (120 kDa), hZP3 (58 kDa) and hZP4 (65 kDa) were purified from ZP (purity >88) by immunoaffinity columns. The binding sites of the purified native hZP3 and hZP4 were localized to the acrosome region of the capacitated human spermatozoa, and were lost after acrosome reaction. Purified human hZP2 bound to this region only in acrosome-reacted spermatozoa. Differential binding of the three glycoproteins to the post-acrosomal region and the midpiece of the spermatozoa was observed. In addition, hZP3, but not hZP2 and hZP4, induced hyperactivation. The stimulatory activity was dependent partly on N-linked glycosylation of hZP3.

CONCLUSIONS

This manuscript describes the biological activities of purified hZP glycoproteins from the native source for the first time.

Keywords: zona pellucida; spermatozoa; glycoprotein; antibody; acrosome

Document Type: Research article

DOI: http://dx.doi.org/10.1093/humrep/den047

Publication date: 2008-06-21

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Human Reproduction features full-length, peer-reviewed papers reporting original research, clinical case histories, as well as opinions and debates on topical issues. Papers published cover the scientific and medical aspects of reproductive physiology and pathology, endocrinology, andrology, gonad function, gametogenesis, fertilization, embryo development, implantation, pregnancy, genetics, genetic diagnosis, oncology, infectious disease, surgery, contraception, infertility treatment, psychology, ethics and social issues. The highest scientific and editorial standard is maintained throughout the journal along with a rapid rate of publication.

Native human zona pellucida glycoproteins: purification and binding properties

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