Phosphatase active antigens in sea urchin eggs and embryos. I. Substrate specificity, pH-optima and inhibitors.
Phosphatase activity in sea urchin embryonic antigens was investigated by histochemical staining of immunoprecipitates separated by two-dimensional (crossed) immunoelectrophoresis. Unfertilized eggs were homogenized in a hypotonic medium which solubilized cytoplasmic antigens. Antigens integrated in membranes or enclosed in particles were solubilized by detergent treatment of the residual pellet. Two different phosphatase activities were discerned in the unfertilized eggs, nucleoside diphosphatase (EC 3.6.1.6.) and acid phosphatase (EC 3.1.3.2.). Nucleoside diphosphatase activity was obtained in both the water soluble and detergent extracted protein fractions. This activity was confined to one antigen. Acid phosphatase acitivity on the other hand was almost exclusively obtained in the detergent extracted fraction and about ten distinct antigens displayed this activity. The nucleoside diphosphatase active antigen preferentially hydrolyzed purine nucleoside diphosphates and to a lesser degree triphosphates of these nucleosides. The acid phosphatase active antigens had a broader substrate specificity and hydrolyzed equally well beta-glycerophosphate and nucleotides. Both activities were essentially inactive at neutral or alkaline pH values. The activities were inhibited by p-choloromercuribenzoate and accordingly stimulated by cysteine. Tartrate and sodium fluoride, however, inhibited the acid phosphatase activity while nucleoside diphosphatase activity was either stimulated or not affected at all by these agents.
D000135:Acid Phosphatase / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D000818:Animals; D000941:Antigens / Q000302:isolation & purification; D002729:Chloromercuribenzoates / Q000494:pharmacology; D003545:Cysteine / Q000494:pharmacology; D004789:Enzyme Activation; D005260:Female; D005306:Fertilization; D005459:Fluorides / Q000494:pharmacology; D005994:Glycerophosphates; D006863:Hydrogen-Ion Concentration; D007122:Immunoelectrophoresis; D009711:Nucleotides; D010063:Ovum / Q000201:enzymology / Q000276:immunology; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D011684:Purine Nucleosides; D012617:Sea Urchins / Q000196:embryology / Q000201:enzymology / Q000276:immunology*; D013644:Tartrates / Q000494:pharmacology
D000941:Antigens; D002729:Chloromercuribenzoates; D005994:Glycerophosphates; D009711:Nucleotides; D011684:Purine Nucleosides; D013644:Tartrates; D000135:Acid Phosphatase; D010744:Phosphoric Monoester Hydrolases; D003545:Cysteine; D005459:Fluorides