pH-dependent cold lability of rabbit skeletal muscle AMP deaminase

Abstract

Rabbit skeletal muscle AMP deaminase (AMP aminohydrolase, EC 3.5.4.6) at low temperature and pH value above 7 undergoes inactivation, with a half-time of the order of several minutes. The loss of activity becomes more extensive at lower enzyme concentrations and higher pH values. It is reversible, since cold-inactivated AMP deaminase can be reactivated by raising the temperature, but not by lowering the pH, of the incubation mixture. The residual activity at the end of the inactivation process at various temperatures, reflecting the equilibrium between active and inactive forms of the enzyme, has been studied as a function of pH to determine the apparent pK and heat of ionization of the process. A general mechanism of reversible inactivation of AMP deaminase is postulated which assumes that deprotonation of the enzyme is followed by isomerization to a form which at low temperature slowly dissociates into the less-active subunits. Cold-inactivated AMP deaminase no longer shows the pH-dependent sigmoidal behaviour of the native enzyme, but regains this property along with the reactivation process. This suggests that allosteric kinetics at basic pH are probably produced by the same isomerization process which is involved in the mechanism for cold lability of the enzyme.