title: Characterization of a partially purified adenosine triphosphatase from a corn root plasma membrane fraction.
abstract: The (K(+),Mg(2+))-ATPase was partially purified from a plasma membrane fraction from corn roots (WF9 x Mol7) and stored in liquid N(2) without loss of activity. Specific activity was increased 4-fold over that of the plasma membrane fraction. ATPase activity resembled that of the plasma membrane fraction with certain alterations in cation sensitivity. The enzyme required a divalent cation for activity (Co(2+) > Mg(2+) > Mn(2+) > Zn(2+) > Ca(2+)) when assayed at 3 millimolar ATP and 3 millimolar divalent cation at pH 6.3. When assayed in the presence of 3 millimolar Mg(2+), the enzyme was further activated by monovalent cations (K(+), NH(4) (+), Rb(+) >> Na(+), Cs(+), Li(+)). The pH optima were 6.5 and 6.3 in the absence and presence of 50 millimolar KCl, respectively. The enzyme showed simple Michaelis-Menten kinetics for the substrate ATP-Mg, with a K(m) of 1.3 millimolar in the absence and 0.7 millimolar in the presence of 50 millimolar KCl. Stimulation by K(+) approached simple Michaelis-Menten kinetics, with a K(m) of approximately 4 millimolar KCl. ATPase activity was inhibited by sodium orthovanadate. Half-maximal inhibition was at 150 and 35 micromolar in the absence and presence of 50 millimolar KCl. The enzyme required the substrate ATP. The rate of hydrolysis of other substrates, except UDP, IDP, and GDP, was less than 20% of ATP hydrolysis. Nucleoside diphosphatase activity was less than 30% of ATPase activity, was not inhibited by vanadate, was not stimulated by K(+), and preferred Mn(2+) to Mg(2+). The results demonstrate that the (K(+),Mg(2+))-ATPase can be clearly distinguished from nonspecific phosphohydrolase and nucleoside diphosphatase activities of plasma membrane fractions prepared from corn roots.
authors: Section of Plant Biology, Division of Biological Sciences, Plant Science Building, Cornell University, Ithaca, New York 14853.
authors: F M
authors: FM
authors: Dupont
authors: L L
authors: LL
authors: Burke
authors: R M
authors: RM
authors: Spanswick