title: The role of Mg2+ in the hydrolytic activity of the isolated chloroplast ATPase: study by high-performance liquid chromatography.
abstract: The influences of total magnesium ion concentration at different total ATP concentrations, and of total ATP concentration, for different total magnesium ion concentrations, on the enzymatic rate of the isolated chloroplast F1 ATPase, have been followed by a chromatographic method consisting in the separation and determination of ADP. From the various series of curves, it is concluded that the experimental results (position of the maxima, Km values) are better fitted by a mechanism involving the activation of the enzyme by magnesium ion and hydrolysis of free ATP, rather than by the classical mechanism, for which the enzyme hydrolyzes the MgATP complex and is inhibited by Mg2+. Although the equations giving the reaction rate are similar in the two cases, the calculated values of Km are widely different. The value obtained from the classical mechanism does not agree with KD, the dissociation constant of the enzyme-substrate complex, measured by the Hummel and Dreyer method. Moreover, when the total ATP concentration tends toward the total magnesium ion concentration, the nucleotide binding to the enzyme tends toward zero, although it should be maximum if MgATP were the true substrate. Finally, the inhibitory effect of Na+ is more easily explained as a competition between this ion and the activating Mg2+, than by the classical mechanism.
authors.affiliation: Département de Biologie Cellulaire et Moléculaire, CEN Saclay, Gif sur Yvette, France.
authors.forename: G
authors.initials: G
authors.lastname: Berger
authors.forename: G
authors.initials: G
authors.lastname: Girault
authors.forename: J M
authors.initials: JM
authors.lastname: Galmiche
authors.forename: S
authors.initials: S
authors.lastname: Pezennec
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D008274:Magnesium / Q000378:metabolism*; D008433:Mathematics; D006180:Proton-Translocating ATPases / Q000302:isolation & purification / Q000378:metabolism*; D012964:Sodium / Q000494:pharmacology
chemical_list: D000255:Adenosine Triphosphate; D012964:Sodium; D006180:Proton-Translocating ATPases; D008274:Magnesium