Document Type
Article
Original Publication Date
2011
Journal/Book/Conference Title
PLOS ONE
Volume
6
DOI of Original Publication
10.1371/journal.pone.0025813
Date of Submission
November 2014
Abstract
The type 1 skeletal muscle ryanodine receptor (RyR1) is principally responsible for Ca2+release from the sarcoplasmic reticulum and for the subsequent muscle contraction. The RyR1 contains three SPRY domains. SPRY domains are generally known to mediate protein-protein interactions, however the location of the three SPRY domains in the 3D structure of the RyR1 is not known. Combining immunolabeling and single-particle cryo-electron microscopy we have mapped the SPRY2 domain (S1085-V1208) in the 3D structure of RyR1 using three different antibodies against the SPRY2 domain. Two obstacles for the image processing procedure; limited amount of data and signal dilution introduced by the multiple orientations of the antibody bound in the tetrameric RyR1, were overcome by modifying the 3D reconstruction scheme. This approach enabled us to ascertain that the three antibodies bind to the same region, to obtain a 3D reconstruction of RyR1 with the antibody bound, and to map SPRY2 to the periphery of the cytoplasmic domain of RyR1. We report here the first 3D localization of a SPRY2 domain in any known RyR isoform.
Rights
Copyright: © 2011 Perálvarez-Marín et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Is Part Of
VCU Physiology and Biophysics Publications
Complete set of identified RyR1 single-particles incubated with anti-SPRY2 antibodies presenting additional mass (indicated by white arrows).
Comments
Originally Published at http://dx.doi.org/10.1371/journal.pone.0025813