Studies on (Na+ + K+)-activated ATPase. XLIV. Single phosphate incorporation during dual phosphorylation by inorganic phosphate and adenosine triphosphate.
(Na+ + K+)-ATPase can be phosphorylated by its substrate ATP as well as by its product inorganic phosphate. The maximal capacity for phosphorylation by either of these two substances is one mol phosphate per mol enzyme. In order to investigate whether the enzyme molecule possesses only on phosphorylation site common to ATP and Pi, or two phosphorylation sites, one for ATP and one for Pi, dual phosphorylation of the enzyme has been carried out. Under conditions, which are maximally favourable for each type of phosphorylation, successive phosphorylation by Pi and ATP leads to a maximal incorporation of only one mol phosphate per mol enzyme. The phosphorylation capacity for ATP decreases by the same amount as the Pi-phosphorylation level increases, without an effect on the apparent affinity for ATP. The results can be explained by assuming either a single common phosphorylation site for Pi and ATP, or a conformational change of the enzyme following phosphorylation by Pi, which excludes phosphorylation by ATP.
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D007679:Kidney Medulla / Q000201:enzymology*; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D010710:Phosphates / Q000378:metabolism*; D010766:Phosphorylation; D011817:Rabbits; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
D010710:Phosphates; D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase