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Isolation, characterization, and localization of the zona pellucida binding proteins of boar sperm
Author(s) -
Hanqing Mo,
TaiYing Yan,
ZhaoWen Shen
Publication year - 1991
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080280204
Subject(s) - zona pellucida , boar , biology , sperm , oocyte , affinity chromatography , microbiology and biotechnology , sepharose , biochemistry , genetics , embryo , enzyme
In order to isolate the putative zona pellucida‐binding proteins (ZPBPs) from boar spermatozoa, a new, simple method has been developed. The new isolation strategy made the most of the highly specific interactions between the components of the gametes. Detergent‐extracted boar sperm proteins were submitted to affinity chromatography on a ZP‐Sepharose column. SDS‐PAGE analyses of the retained fraction under reducing conditions revealed that in addition to a component of M r 38,000, the predominant ZPBPs contained at least three low molecular weight proteins (M r <20 kDa). The isolated ZPBPs were effective in blocking sperm‐oocyte binding in vitro. Using immunofluorescence microscopy, the ZPBPs were shown to be localized primarily in the sperm head, especially in the acrosomal cap region.