Plasma membrane association, purification, and partial characterization of mouse sperm beta 1,4-galactosyltransferase.
Gamete recognition in the mouse is mediated, in part, by the binding of sperm surface galactosyltransferase (GalTase) to appropriate substrates in the egg zona pellucida. In this paper, sperm GalTase is shown to be an externally oriented, integral plasma membrane component. GalTase is not peripherally adsorbed to the cell surface, nor is it bound to cell surface glycoside substrates. GalTase can be released from the surface of intact sperm by either mild proteolysis or by detergent under conditions in which the sperm membranes remain intact as judged by double-label indirect immunofluorescence. Detergent-solubilized sperm GalTase has been purified to apparent homogeneity by affinity chromatography and characterized as a beta 1,4-GlcNAc:GalTase by substrate and kinetic analyses. Purified and membrane-bound GalTase both show an unusual thermal inactivation above 39-40 degrees C, whereas other sperm enzyme activities as well as GalTase activity from other cell types are temperature-dependent. Purified sperm GalTase inhibits sperm binding to the egg zona pellucida, consistent with its proposed role during gamete recognition.
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002846:Chromatography, Affinity; D005260:Female; D007700:Kinetics; D007788:Lactose Synthase / Q000302:isolation & purification*; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009237:N-Acetyllactosamine Synthase / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000201:enzymology*; D013379:Substrate Specificity
D007788:Lactose Synthase; D009237:N-Acetyllactosamine Synthase