Native human zona pellucida glycoproteins: purification and binding properties

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...

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Bibliographic Details
Published in:Human reproduction (Oxford) 2008-06, Vol.23 (6), p.1385-1393
Main 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.
Format: Article
Language:English
Subjects:
acrosome
Acrosome - metabolism
Antibodies, Monoclonal
antibody
Biological and medical sciences
Egg Proteins - isolation & purification
Egg Proteins - pharmacology
Female
glycoprotein
Gynecology. Andrology. Obstetrics
Humans
Male
Medical sciences
Membrane Glycoproteins - isolation & purification
Membrane Glycoproteins - pharmacology
Receptors, Cell Surface - isolation & purification
spermatozoa
zona pellucida
Zona Pellucida - metabolism
Zona Pellucida Glycoproteins
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Summary: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.
ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/den047