Reversible inactivation of d-fructose 1,6-diphosphatase by adenosine triphosphate and cyclic 3′, 5′-adenosine monophosphate

Abstract

An analytical method for the quantitative recovery and purification of fructose 1,6-diphosphatase from incubation mixtures containing crude kidney extracts has been described. Utilizing this procedure it has been demonstrated that renal fructose 1,6-diphosphatase was inactivated on incubation with ATP and cyclic-3′,5′-AMP. Fructose 1,6-diphosphatase present in crude kidney extracts was inactivated by ATP and Mg ++ alone, and the rate of this inactivation was increased in the presence of cyclic 3′,5′-AMP. Purified enzyme was inactivated by ATP only in the presence of crude kidney extracts. Fructose 1,6-diphosphatase was also inactivated when kidney slices were incubated in the presence of epinephrine. When fructose 1,6-diphosphatase was incubated in the presence of an AT 32P generating system in crude extracts, 32P was incorporated into the enzyme protein with a concomitant decrease in enzyme activity. Very little or no 32P i was incorporated in the absence of adenosine nucleotide. The inactivated enzyme was reactivated on incubation with crude kidney extracts. The possible relationship of these effects to those obtained with three other enzymes involved in the synthesis and degradation of glycogen and the mechanism of action of epinephrine is discussed.