BK channel modulation by positively charged peptides and auxiliary γ subunits mediated by the Ca²⁺-bowl site

The large-conductance, Ca²⁺-, and voltage-activated K⁺ (BK) channel consists of the pore-forming α (BKα) subunit and regulatory β and γ subunits. The γ1–3 subunits facilitate BK channel activation by shifting the voltage-dependence of channel activation toward the hyperpolarization direction by about 50–150 mV in the absence of Ca²⁺. We previously found that the intracellular C-terminal positively charged regions of the γ subunits play important roles in BK channel modulation. In this study, we found that the intracellular C-terminal region of BKα is indispensable in BK channel modulation by the γ1 subunit. Notably, synthetic peptide mimics of the γ1–3 subunits’ C-terminal positively charged regions caused 30–50 mV shifts in BKα channel voltage-gating toward the hyperpolarization direction. The cationic cell–penetrating HIV-1 Tat peptide exerted a similar BK channel–activating effect. The BK channel–activating effects of the synthetic peptides were reduced in the presence of Ca²⁺ and markedly ablated by both charge neutralization of the Ca²⁺-bowl site and high ionic strength, suggesting the involvement of electrostatic interactions. The efficacy of the γ subunits in BK channel modulation was reduced by charge neutralization of the Ca²⁺-bowl site. However, BK channel modulation by the γ1 subunit was little affected by high ionic strength and the positively charged peptide remained effective in BK channel modulation in the presence of the γ1 subunit. These findings identify positively charged peptides as BK channel modulators and reveal a role for the Ca²⁺-bowl site in BK channel modulation by positively charged peptides and the C-terminal positively charged regions of auxiliary γ subunits.