Purification of sea urchin sperm bindin by DEAE-cellulose chromatography.
A procedure for purifying bindin from sperm of the sea urchin Strongylocentrotus purpuratus is presented in detail. The impure bindin, dissolved in 4 M urea, 50 mM sodium phosphate, pH 6.6, is adsorbed to DEAE-cellulose and eluted wit 4 M urea, 650 mM sodium phosphate, pH 6.6. The purified bindin is not contaminated with tubulin or histone HI. A precipitate of this DEAE-purified bindin, made by dialysis into Ca2+-free seawater and natural seawater, is a species-specific agglutinin of unfertilized eggs. This method of obtaining consistently pure preparations of bindin will aid in the analysis of its role in fertilization.
D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification* / Q000494:pharmacology; D008297:Male; D011956:Receptors, Cell Surface; D012617:Sea Urchins; D013084:Sperm-Ovum Interactions / Q000187:drug effects; D013094:Spermatozoa / Q000032:analysis*
D006023:Glycoproteins; D011956:Receptors, Cell Surface; C035027:egg surface sperm receptor