Activation of protein kinase A partially reverses the effects of 2,3-butanedione monoxime on the transient outward K+ current of rat ventricular myocytes.

The transient outward K+ current (Ito) was assessed in single rat ventricular myocytes with the whole-cell patch-clamp technique. Extracellular application of the chemical phosphatase 2,3-butanedione monoxime (BDM) inhibited Ito in a concentration-dependent manner. The IC50 was 14 mM. The on-set of this effect occurred within 20 s after BDM application. Ito recovered almost completely at 2 min after washout of BDM. Application of 20 mM BDM shifted the steady-state inactivation curve of Ito by 9 +/- 2 mV (n = 8) in the negative direction. Addition of 5 microM isoproterenol enhanced Ito amplitude by 16.2 +/- 1.8%. This concentration of isoproterenol partially reversed the BDM-induced inhibition of Ito. Furthermore, application of 10 mM 8-bromoadenosine 3':5'-cyclic monophosphate enhanced the amplitude of Ito and also significantly reversed the BDM-induced suppression of Ito. In contrast, intracellular dialysis with guanosine 3':5'-cyclic monophosphate (cGMP, 1-10 mM) did not affect the BDM-induced inhibition of Ito. The inward rectifier K+ current (Ik1) was relatively insensitive to BDM; i.e., 20 mM BDM inhibited Ito and Ik1 to 35.5 +/- 4.3% (n = 8) and 92.9 +/- 4.0% (n = 4) of the control, respectively. These results indicate that BDM suppressed Ito but not Ik1 of rat ventricular myocytes. We attribute the BDM suppression of Ito to dephosphorylation of the channel protein.