Polyunsaturated fatty acids inhibit Kv1.4 by interacting with positively charged extracellular pore residues

Polyunsaturated fatty acids (PUFAs) modulate voltage-gated K+ channel inactivation by an unknown site and mechanism. The effects of omega-6 and omega-3 PUFAs were investigated on the heterologously expressed K(v)1.4 channel. PUFAs inhibited wild-type K(v)1.4 during repetitive pulsing as a result of slowing of recovery from inactivation. In a mutant K(v)1.4 channel lacking N-type inactivation, PUFAs reversibly enhanced C-type inactivation (Kd, 15-43 mu M). C-type inactivation was affected by extracellular H+ and K+ as well as PUFAs and there was an interaction among the three: the effect of PUFAs was reversed during acidosis and abolished on raising K+. Replacement of two positively charged residues in the extracellular pore (H508 and K532) abolished the effects of the PUFAs (and extracellular H+ and K+) on C-type inactivation but had no effect on the lipoelectric modulation of voltage sensor activation, suggesting two separable interaction sites/mechanisms of action of PUFAs. Charge calculations suggest that the acidic head group of the PUFAs raises the pK(a) of H508 and this reduces the K+ occupancy of the selectivity filter, stabilizing the C-type inactivated state.