Calyculin A and outward K+ channel currents in rat tail artery smooth muscle cells

Protein dephosphorylation mediated by phosphatases represents an important mechanism for modulating the functions of the targeted proteins. Calyculin A has been extensively used as a specific inhibitor of protein phosphatases. However, the effect of calyculin A on K channel currents in vascular smooth muscle cells (SMCs) and the underlying mechanisms had been unknown. It was found in the current study that calyculin A inhibited the whole-cell outward K channel currents in rat tail artery SMCs in a concentration-dependent (median inhibitory concentration, 12.6 n ) and reversible fashion. The extracellular applied calyculin A induced a biphasic change in K current amplitude with an initial transient increase followed by a long-lasting inhibition (n = 6). The intracellularly applied calyculin A (100 nM ) caused a lesser inhibition (33 +/- 1%) of K channel currents than that caused by the extracellularly applied calyculin A (55.3 +/- 8% inhibition) and did not result in an initial increase in K channel currents. The inhibitory effect of the intracellularly applied calyculin A on K channel currents was reversed to a stimulatory effect after ATP was omitted from the intracellular solution. The K currents inhibited by calyculin A were conducted by the iberiotoxin-sensitive K channels in SMCs. Moreover, okadaic acid (0.03-3 microM ) did not cause any significant change in K(Ca) channel currents. In conclusion, calyculin A inhibited K(Ca) channel currents in vascular SMCs. This effect of calyculin A, however, was not mediated by the inhibition of protein phosphatases.