Mechanistic investigation on the temperature dependence and inhibition of corn root plasma membrane ATPase.
The kinetics of corn root plasma membrane-catalyzed Mg-ATP hydrolysis may be satisfactorily described by a simple Michaelis-Menten scheme. It was found that the Km of the process was relatively insensitive to changes in temperature. This property allowed us to conveniently estimate the activation energy of the enzyme turnover process as approximately 14 kcal mol-1 in the temperature range of 10 to 45 degrees C. The enzyme activity was inhibited by the presence of diethystilbestrol (DES), miconazole, vanadate, and dicyclohexylcarbodiimide (DCCD). The inhibition caused by DES and miconazole was strictly uncompetitive and inhibition by vanadate was noncompetitive. The inhibition by DCCD showed a substrate concentration dependence, i.e., competitive at high and uncompetitive at low concentrations of Mg-ATP. The 1/V vs [I] plots suggested that there were different but unique binding sites for DES, vanadate, and miconazole. However, the modification of the plasma membrane by DCCD exhibited interaction with multiple sites. Unlike yeast plasma membrane ATPase, the enzyme of corn root cells was not affected by the treatment with N-ethylmaleimide. Although the enzyme activity was regulated by ADP, a product of the reaction, the presence of inorganic phosphate showed no inhibition to the hydrolysis of Mg-ATP.
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D000251:Adenosine Triphosphatases / Q000032:analysis* / Q000037:antagonists & inhibitors; D002462:Cell Membrane / Q000201:enzymology; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004054:Diethylstilbestrol / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D007700:Kinetics; D008825:Miconazole / Q000494:pharmacology; D010710:Phosphates / Q000494:pharmacology; D013696:Temperature; D014638:Vanadates; D014639:Vanadium / Q000494:pharmacology; D003313:Zea mays / Q000201:enzymology*
D010710:Phosphates; D014639:Vanadium; D014638:Vanadates; D004024:Dicyclohexylcarbodiimide; D004054:Diethylstilbestrol; D008825:Miconazole; D000251:Adenosine Triphosphatases; D005033:Ethylmaleimide