Molecular mechanisms for large conductance Ca2+-activated K+ channel activation by a novel opener, 12,14-dichlorodehydroabietic acid

Our recent study has revealed that 12,14-dichlorodehydroabietic acid (diCl-DHAA), which is synthetically derived from a natural product, abietic acid, is a potent opener of large conductance CA(2+)-activated K(+) (BK) channel. Here, we examined, by using a channel expression system in human embryonic kidney 293 cells, the mechanisms underlying the BK channel opening action of diCl-DHAA and which subunit of the BK channel (alpha or beta1) is the site of action for diCl-DHAA. BK channel activity was significantly enhanced by diCl-DHAA at concentrations of 0.1 microM and higher in a concentration-dependent manner. diCl-DHAA enhanced the activity of BKalpha by increasing sensitivity to both CA(2+) and membrane potential without changing the single channel conductance. It is notable that the increase in BK channel open probability by diCl-DHAA showed significant inverse voltage dependence, i.e., larger potentiation at lower potentials. Since coexpression of beta1 subunit with BKalpha did not affect the potency of diCl-DHAA, the site of action for diCl-DHAA is suggested to be BKalpha subunit. Moreover, kinetic analysis of single channel currents indicates that diCl-DHAA opens BKalpha mainly by decreasing the time staying in a long closed state. Although reconstituted voltage-dependent CA(2+) channel current was significantly reduced by 1 microM diCl-DHAA, BK channels were selectively activated at lower concentrations. These results indicate that diCl-DHAA is one of the most potent BK channel openers acting on BKalpha and a useful prototype compound to develop a novel BK channel opener.