Isolation and characterization of a plasma membrane fraction from sea urchin sperm exhibiting species specific recognition of the egg surface.
A method is described for isolating preparative quantities of plasma membranes from sea urchin sperm. The final membrane fraction is homogeneous by sucrose density sedimentation and is enriched in adenylate cyclase as well as in the four glycoproteins accessible to radioiodination of intact sperm. The electrophoretic profiles of sperm membranes from three sea urchin species are very similar. The membrane preparation consists primarily of sealed vesicles which release carboxyfluorescein when exposed to detergents or distilled water. Ninety-two percent of the 125I-labeled vesicle material binds to wheat germ lectin columns, suggesting a right-side-out orientation. The isolated sperm membrane vesicles exhibit species specific adhesion to the surfaces of sea urchin eggs; this adhesion is blocked by pretreatment of the vesicles with trypsin or egg jelly. This method will be useful for isolating biologically active sperm membrane components involved in sperm-egg recognition during fertilization.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000502:physiology* / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D003576:Electron Transport Complex IV / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification; D008854:Microscopy, Electron; D008970:Molecular Weight; D012617:Sea Urchins / Q000502:physiology*; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000648:ultrastructure*
D005452:Fluoresceins; D005456:Fluorescent Dyes; D008565:Membrane Proteins; C024098:6-carboxyfluorescein; D003576:Electron Transport Complex IV