3-D Structural and Functional Characterization of the Purified KATP Channel Complex Kir6.2-SUR1

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2005 Nov 26

PMID 16308567

Citations 80

Authors

Michael V Mikhailov

Jeff D Campbell

Heidi de Wet

Kenju Shimomura

Brittany Zadek

Richard F Collins

Mark S P Sansom

Robert C Ford

Frances M Ashcroft

Affiliations

Soon will be listed here.

Abstract

ATP-sensitive potassium (K(ATP)) channels conduct potassium ions across cell membranes and thereby couple cellular energy metabolism to membrane electrical activity. Here, we report the heterologous expression and purification of a functionally active K(ATP) channel complex composed of pore-forming Kir6.2 and regulatory SUR1 subunits, and determination of its structure at 18 A resolution by single-particle electron microscopy. The purified channel shows ATP-ase activity similar to that of ATP-binding cassette proteins related to SUR1, and supports Rb(+) fluxes when reconstituted into liposomes. It has a compact structure, with four SUR1 subunits embracing a central Kir6.2 tetramer in both transmembrane and cytosolic domains. A cleft between adjacent SUR1s provides a route by which ATP may access its binding site on Kir6.2. The nucleotide-binding domains of adjacent SUR1 appear to interact, and form a large docking platform for cytosolic proteins. The structure, in combination with molecular modelling, suggests how SUR1 interacts with Kir6.2.

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