Structure of an open KATPchannel reveals tandem PIP2binding sites mediating the Kir6.2 and SUR1 regulatory interface

Abstract

AbstractATP-sensitive potassium (KATP) channels, composed of four pore-lining Kir6.2 subunits and four regulatory sulfonylurea receptor 1 (SUR1) subunits, control insulin secretion in pancreatic β-cells. KATPchannel opening is stimulated by PIP2and inhibited by ATP. Mutations that increase channel opening by PIP2reduce ATP inhibition and cause neonatal diabetes. Although considerable evidence has implicated a role for PIP2in KATPchannel function, previously solved open-channel structures have lacked bound PIP2, and mechanisms by which PIP2regulates KATPchannels remain unresolved. Here, we report cryoEM structure of a KATPchannel harboring the neonatal diabetes mutation Kir6.2-Q52R, bound to natural C18:0/C20:4 long-chain PI(4,5)P2in open conformation. The structure reveals two adjacent PIP2molecules between SUR1 and Kir6.2. The first PIP2binding site is conserved among PIP2-gated Kir channels. The second site forms uniquely in KATPat the interface of Kir6.2 and SUR1. Functional studies demonstrate both binding sites determine channel activity. Kir6.2 pore opening is associated with a twist of the Kir6.2 cytoplasmic domain and a rotation of the N-terminal transmembrane domain of SUR1, which widens the inhibitory ATP binding pocket to disfavor ATP binding. The open conformation is particularly stabilized by the Kir6.2-Q52R residue through cation-π bonding with SUR1 - W51. Together, these results uncover the cooperation between SUR1 and Kir6.2 in PIP2binding and gating, explain the antagonistic regulation of KATPchannels by PIP2and ATP, and provide the mechanism by which Kir6.2-Q52R stabilizes an open channel to cause neonatal diabetes.