Selective Activation of Calcineurin by Dipicolinic Acid

Calcineurin was activated at 30°C by incubation with dipicolinic acid, a metal chelator, in the absence of activating, exogenous Mn2+. The activation reached a plateau after 90 min with 8- to 12-fold higher activity. Inclusion of the activating metal Mn2+(1.0 mm) in the incubation mixture slightly lessened the activation induced by dipicolinic acid. The chelator 1,10-phenanthroline had no effect on the activity of calcineurin in concurrent experiments. Activation by dipicolinic acid was reversed by the addition of Zn2+or Fe3+. The reversal occurred within 30 min after the addition of either metal and returned the activity of calcineurin to its initial level. Atomic absorption spectrometry analysis showed no loss of iron or zinc from calcineurin after activation (2 h) by dipicolinic acid. Because there seemed to be no interaction between dipicolinic acid and exogenous metal, the effect of dipicolinic acid was concluded to result from masking of at least one intrinsic metal. Calcineurin incubated with 1.0 mmMn2+(saturating levels) also did not show any loss of intrinsic metal by atomic absorption analysis. The consequences of these data concerning the role(s) of intrinsic metals in calcineurin catalysis are discussed.