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Single-particle cryo-EM of the ryanodine receptor channel

Mariah R. Baker, Guizhen Fan, Irina I. Serysheva
  • Mariah R. Baker
    Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, TX, United States
  • Guizhen Fan
    Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, TX, United States
  • Irina I. Serysheva
    Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, TX, United States | irina.i.serysheva@uth.tmc.edu

Abstract

Ryanodine receptors (RyRs) are tetrameric ligand-gated Ca2+ release channels that are responsible for the increase of cytosolic Ca2+ concentration leading to muscle contraction. Our current understanding of RyR channel gating and regulation is greatly limited due to the lack of a high-resolution structure of the channel protein. The enormous size and unwieldy shape of Ca2+ release channels make X-ray or NMR methods difficult to apply for high-resolution structural analysis of the full-length functional channel. Single-particle electron cryo-microscopy (cryo-EM) is one of the only effective techniques for the study of such a large integral membrane protein and its molecular interactions. Despite recent developments in cryo-EM technologies and break-through single-particle cryo-EM studies of ion channels, cryospecimen preparation, particularly the presence of detergent in the buffer, remains the main impediment to obtaining atomic-resolution structures of ion channels and a multitude of other integral membrane protein complexes. In this review we will discuss properties of several detergents that have been successfully utilized in cryo-EM studies of ion channels and the emergence of the detergent alternative amphipol to stabilize ion channels for structure-function characterization. Future structural studies of challenging specimen like ion channels are likely to be facilitated by cryo-EM amenable detergents or alternative surfactants.

Keywords

Ryanodine receptor, Membrane proteins, Electron cryo-microscopy, Cryospecimen preparation, Detergents, Amphipol

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Submitted: 2014-10-29 13:11:45
Published: 2015-01-12 09:11:45
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Copyright (c) 2015 Mariah R. Baker, Guizhen Fan, Irina I. Serysheva

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