Genistein and tyrphostin AG556 decrease ultra-rapidly activating delayed rectifier K⁺ current of human atria by...

The ultra-rapidly activating delayed rectifier K⁺ current I_Kur (encoded by K_v 1.5 or KCNA5) plays an important role in human atrial repolarization. The present study investigates the regulation of this current by protein tyrosine kinases (PTKs).

Whole-cell patch voltage clamp technique and immunoprecipitation and Western blotting analysis were used to investigate whether the PTK inhibitors genistein, tyrphostin AG556 (AG556) and PP2 regulate human atrial I_Kur and hKv1.5 channels stably expressed in HEK 293 cells.

Human atrial I_Kur was decreased by genistein (a broad-spectrum PTK inhibitor) and AG556 (a highly selective EGFR TK inhibitor) in a concentration-dependent manner. Inhibition of I_Kur induced by 30 μM genistein or 10 μM AG556 was significantly reversed by 1 mM orthovanadate (a protein tyrosine phosphatase inhibitor). Similar results were observed in HEK 293 cells stably expressing hK_v 1.5 channels. On the other hand, the Src family kinase inhibitor PP2 (1 μM) slightly enhanced I_Kur and hK_v 1.5 current, and the current increase was also reversed by orthovanadate. Immunoprecipitation and Western blotting analysis showed that genistein, AG556, and PP2 decreased tyrosine phosphorylation of hK_v 1.5 channels and that the decrease was countered by orthovanadate.

The PTK inhibitors genistein and AG556 decrease human atrial I_Kur and cloned hK_v 1.5 channels by inhibiting EGFR TK, whereas the Src kinase inhibitor PP2 increases I_Kur and hK_v 1.5 current. These results imply that EGFR TK and the soluble Src kinases may have opposite effects on human atrial I_Kur .