Isolation, characterization, and localization of the zona pellucida binding proteins of boar sperm.
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 Mr 38,000, the predominant ZPBPs contained at least three low molecular weight proteins (Mr less than 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.
Mo
H Q
HQ
Glycoconjugate and Lectin Research Laboratory, Shanghai Institute of Biochemistry, Academia Sinica, China.
Yan
T Y
TY
Shen
Z W
ZW
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002469:Cell Separation; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005455:Fluorescent Antibody Technique; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000737:chemistry* / Q000378:metabolism; D013552:Swine; D015044:Zona Pellucida / Q000378:metabolism*
D008565:Membrane Proteins