A Mg2+- and Ca2+-stimulated adenosine triphosphatase at the outer surface of Ehrlich ascites tumor cells.
A Mg2+- and Ca2+-stimulated adenosine triphosphatase (ATPase) at the outer surface of intact Ehrlich ascites tumor cells is described. A surface-bound adenosine triphosphate (ATP)-splitting activity at a lower rate was also demonstrated in the absence of Ca2+ but with Mg2+, Na+, and K+ present in the isotonic medium. Hence, when part of the Mg2+ was exchanged for Ca2+, a marked increase of the ATP-splitting activity was observed. The stimulatory effect of Ca2+ was seen only if both Na+ and K+ were present in the isotonic incubation medium. Thus, the enzyme activity was Mg2+- and Ca2+-dependent. Ca2+, together with the monovalent cations was inhibitory compared with Mg2+ under similar conditions. The apparent Km for ATP for the Mg2+-stimulated ATPase is 0.05 mM, while that of the Mg2+- and Ca2+-stimulated enzyme is 0.10 mM. The Vmax of the former is 0.8 mu-mole per 100 mg Schneider protein per 30 sec compared with 1.92 mu-moles per 100 mg Schneider protein per 30 sec for the latter. The calculated Km for the Mg2+- and Ca2+-stimulated ATPase after subtraction of the Mg2+-stimulated part is 0.22 mM. Ethacrynic acid and N-ethylmaleimide both inhibited the Mg2+- and Ca2+-stimulated ATPase by about 10 percent, while the ouabain inhibition was 15 percent. Cytochalasin B did not influence the enzyme activity, whereas La3+ had a slight stimulatory effect.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D003571:Cytochalasin B / Q000494:pharmacology; D004789:Enzyme Activation; D004976:Ethacrynic Acid / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D066298:In Vitro Techniques; D007811:Lanthanum / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D051379:Mice; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D012964:Sodium / Q000494:pharmacology; D013268:Stimulation, Chemical
D003571:Cytochalasin B; D010042:Ouabain; D007811:Lanthanum; D000255:Adenosine Triphosphate; D012964:Sodium; D000251:Adenosine Triphosphatases; D008274:Magnesium; D004976:Ethacrynic Acid; D005033:Ethylmaleimide; D011188:Potassium; D002118:Calcium