Structural basis of human Slo2.2 channel gating and modulation

Abstract

The sodium-activated Slo2.2 channel is abundantly expressed in the brain, playing a critical role in regulating neuronal excitability. The Na⁺-binding site and the underlying mechanisms of Na⁺-dependent activation remain unclear. Here, we present cryoelectron microscopy (cryo-EM) structures of human Slo2.2 in closed, open, and inhibitor-bound form at resolutions of 2.6-3.2 Å, revealing gating mechanisms of Slo2.2 regulation by cations and a potent inhibitor. The cytoplasmic gating ring domain of the closed Slo2.2 harbors multiple K⁺ and Zn²⁺ sites, which stabilize the channel in the closed conformation. The open Slo2.2 structure reveals at least two Na⁺-sensitive sites where Na⁺ binding induces expansion and rotation of the gating ring that opens the inner gate. Furthermore, a potent inhibitor wedges into a pocket formed by pore helix and S6 helix and blocks the pore. Together, our results provide a comprehensive structural framework for the investigation of Slo2.2 channel gating, Na⁺ sensation, and inhibition.

Keywords: CP: Molecular biology; KCNT1; Slo2.2; cryo-EM; epilepsy; potassium channel; small-molecule inhibition; sodium-dependent activation.