Calcium-stimulated ATPase activity in homogenates of the secretory enamel organ in the rat.
The activity of calcium-stimulated ATPase (E.C. 3.6.1.3) in homogenates of the secretory enamel organ of rat incisors was studied biochemically. ATP hydrolysis was estimated from the amount of inorganic phosphate liberated. An analysis of the total degradation of ATP was initially performed to ensure that the enzyme assays pertained to the original substrate, ATP, and were not influenced by reaction products formed. Standard incubations were run in tris-maleate buffer, pH 8.2, with 3 mM ATP, 3 mM Ca2+ and 0.5 mM R 8231 at 37 degrees C. The presence of R 8231 was necessary to inhibit nonspecific alkaline phosphatase. The calcium-stimulated ATPase was completely inhibited when heated at 55-60 degrees C for 5 min. The pH optimum was found to be 8.2. The hydrolysis was substantially dependent on Ca2+ and was fastest when the ATP:Ca2+ ratio was 1:1. High substrate concentrations inhibited the hydrolysis. The addition of 1 mM Zn2+ and Ni2+ to the incubation medium markedly inhibited the hydrolysis as did, though less strongly, p-hydroxymercuribenzoate, oligomycin, EDTA and ruthenium red. l-Cysteine, mercaptoethanol, iodoacetic acid and sodium azide were without effect. F- was without effect unless added to a final concentration above 15 mM to media where Ca2+ had first been allowed to react with ATP.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D004658:Enamel Organ / Q000201:enzymology*; D005260:Female; D005459:Fluorides / Q000494:pharmacology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D051381:Rats; D013773:Tetramisole / Q000031:analogs & derivatives / Q000494:pharmacology; D014083:Tooth Germ / Q000201:enzymology*
D000255:Adenosine Triphosphate; D013773:Tetramisole; D000251:Adenosine Triphosphatases; D005459:Fluorides; D002118:Calcium