High-resolution structure of the membrane-embedded skeletal muscle ryanodine receptor

Abstract

The type 1 ryanodine receptor (RyR)/calcium release channel on the sarcoplasmic reticulum (SR) is required for skeletal muscle excitation-contraction coupling and is the largest known ion channel, composed of four 565-kDa protomers. Cryogenic electron microscopy (cryo-EM) studies of the RyR have primarily used detergent to solubilize the channel; in the present study, we have used cryo-EM to solve high-resolution structures of the channel in liposomes using a gel-filtration approach with on-column detergent removal to form liposomes and incorporate the channel simultaneously. This allowed us to resolve the structure of the channel in the primed and open states at 3.4 and 4.0 Å, respectively, with a single dataset. This method offers validation for detergent-based structures of the RyR and offers a starting point for utilizing a chemical gradient mimicking the SR, where Ca²⁺ concentrations are millimolar in the lumen and nanomolar in the cytosol.

Keywords: calcium release channel; cryoelectron microscopy; excitation-contraction coupling; ion channel; membrane protein; proteoliposomes; ryanodine receptor; sarcoplasmic reticulum; skeletal muscle.