Secreted epididymal glycoprotein 2D6 that binds to the sperm's plasma membrane is a member of the beta-defensin superfamily of pore-forming glycopeptides.
The plasma membrane of spermatozoa undergoes substantial remodeling during passage through the epididymal duct, principally because of changes in phospholipid composition, exchange of glycoproteins with epididymal fluid, and processing of existing membrane proteins. Here, we describe the interaction of an epididymal glycoprotein recognized by monoclonal antibody 2D6 with the plasma membrane of rat spermatozoa. Our goals have been to understand more about the mechanism of secretion of epididymal glycoproteins, how they interact with the sperm's plasma membrane, and their disposition within it. Reactivity to 2D6 monoclonal antibody was first detectable in principal cells in the distal caput epididymidis and as a soluble high-molecular-weight complex in the secreted fluid. It was not associated with membranous vesicles in the duct lumen. On cauda spermatozoa 2D6 monoclonal antibody recognized a 24-kDa glycoprotein (the subunit of a disulfide cross-linked homodimer of 48 kDa) that was present on the plasma membrane overlying the sperm tail. Binding of 2D6 to immature spermatozoa in vitro was cell-type specific but not species specific, and the antigen could only be extracted from cauda spermatozoa with detergents. Sequencing studies revealed that the 24-kDa glycoprotein was a member of the beta-defensin superfamily of small pore-forming glycopeptides of which several others (ESP13.2, Bin1b, E-2, EP2, HE2) are found in the epididymis. This evidence suggests that some epididymal glycoproteins are secreted into the luminal fluid in a soluble form and bind to specific regions of the sperm's surface via hydrophobic interactions. Given the antimicrobial function of beta-defensins, they have a putative role in protecting spermatozoa and the epididymis from bacterial infections.
Zanich
Alan
A
Gamete Signalling Laboratory, The Babraham Institute, Cambridge CB2 4AT, United Kingdom.
Pascall
John C
JC
Jones
Roy
R
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D004822:Epididymis / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology; D006020:Glycopeptides / Q000378:metabolism; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013094:Spermatozoa / Q000378:metabolism*; D023083:beta-Defensins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D006020:Glycopeptides; D006023:Glycoproteins; D023083:beta-Defensins