Using cryo-electron microscopy and single particle image processing
techniques, we present the first three-dimensional reconstructions
of isoform 3 of the ryanodine receptor/calcium release channel (RyR3).
Reconstructions were carried out on images obtained from a purified,
detergent-solubilized receptor for two different buffer conditions,
which were expected to favor open and closed functional states of
the channel. As for the heart (RyR2) and skeletal muscle (RyR1) receptor
isoforms, RyR3 is a homotetrameric complex comprising two main components,
a multidomain cytoplasmic assembly and a smaller ( approximately
20\% of the total mass) transmembrane region. Although the isoforms
show structural similarities, consistent with the approximately 70\%
overall sequence identity of the isoforms, detailed comparisons of
RyR3 with RyR1 showed one region of highly significant difference
between them. This difference indicated additional mass present in
RyR1, and it likely corresponds to a region of the RyR1 sequence
(residues 1303-1406, known as diversity region 2) that is absent
from RyR3. The reconstructions of RyR3 determined under öpen" and
"closed" conditions were similar to each other in overall architecture.
A difference map computed between the two reconstructions reveals
subtle changes in conformation at several widely dispersed locations
in the receptor, the most prominent of which is a approximately 4
degrees rotation of the transmembrane region with respect to the
cytoplasmic assembly.
%0 Journal Article
%1 Shar_2000_9485
%A Sharma, M. R.
%A Jeyakumar, L. H.
%A Fleischer, S.
%A Wagenknecht, T.
%D 2000
%J J Biol Chem
%K Animals; Buffers; Calcium Cattle; Channel, Conformation; Cryoelectron Isoforms, Microscopy; Models, Molecular; Protein Receptor Release Ryanodine chemistry/ultrastructure chemistry/ultrastructure;
%N 13
%P 9485--9491
%T Three-dimensional structure of ryanodine receptor isoform three in
two conformational states as visualized by cryo-electron microscopy.
%U http://www.jbc.org/cgi/content/full/275/13/9485
%V 275
%X Using cryo-electron microscopy and single particle image processing
techniques, we present the first three-dimensional reconstructions
of isoform 3 of the ryanodine receptor/calcium release channel (RyR3).
Reconstructions were carried out on images obtained from a purified,
detergent-solubilized receptor for two different buffer conditions,
which were expected to favor open and closed functional states of
the channel. As for the heart (RyR2) and skeletal muscle (RyR1) receptor
isoforms, RyR3 is a homotetrameric complex comprising two main components,
a multidomain cytoplasmic assembly and a smaller ( approximately
20\% of the total mass) transmembrane region. Although the isoforms
show structural similarities, consistent with the approximately 70\%
overall sequence identity of the isoforms, detailed comparisons of
RyR3 with RyR1 showed one region of highly significant difference
between them. This difference indicated additional mass present in
RyR1, and it likely corresponds to a region of the RyR1 sequence
(residues 1303-1406, known as diversity region 2) that is absent
from RyR3. The reconstructions of RyR3 determined under öpen" and
"closed" conditions were similar to each other in overall architecture.
A difference map computed between the two reconstructions reveals
subtle changes in conformation at several widely dispersed locations
in the receptor, the most prominent of which is a approximately 4
degrees rotation of the transmembrane region with respect to the
cytoplasmic assembly.
@article{Shar_2000_9485,
abstract = {Using cryo-electron microscopy and single particle image processing
techniques, we present the first three-dimensional reconstructions
of isoform 3 of the ryanodine receptor/calcium release channel (RyR3).
Reconstructions were carried out on images obtained from a purified,
detergent-solubilized receptor for two different buffer conditions,
which were expected to favor open and closed functional states of
the channel. As for the heart (RyR2) and skeletal muscle (RyR1) receptor
isoforms, RyR3 is a homotetrameric complex comprising two main components,
a multidomain cytoplasmic assembly and a smaller ( approximately
20\% of the total mass) transmembrane region. Although the isoforms
show structural similarities, consistent with the approximately 70\%
overall sequence identity of the isoforms, detailed comparisons of
RyR3 with RyR1 showed one region of highly significant difference
between them. This difference indicated additional mass present in
RyR1, and it likely corresponds to a region of the RyR1 sequence
(residues 1303-1406, known as diversity region 2) that is absent
from RyR3. The reconstructions of RyR3 determined under "open" and
"closed" conditions were similar to each other in overall architecture.
A difference map computed between the two reconstructions reveals
subtle changes in conformation at several widely dispersed locations
in the receptor, the most prominent of which is a approximately 4
degrees rotation of the transmembrane region with respect to the
cytoplasmic assembly.},
added-at = {2009-06-03T11:20:58.000+0200},
author = {Sharma, M. R. and Jeyakumar, L. H. and Fleischer, S. and Wagenknecht, T.},
biburl = {https://www.bibsonomy.org/bibtex/21e2844971057a5b1410affc69645c5a2/hake},
description = {The whole bibliography file I use.},
file = {Shar_2000_9485.pdf:Shar_2000_9485.pdf:PDF},
institution = {Wadsworth Center for Laboratories and Research, New York State Department
of Health, Albany, New York 12201-0509, USA. manjuli@wadsworth.org},
interhash = {6f3b5807bc6aad9af3acd7282a640f59},
intrahash = {1e2844971057a5b1410affc69645c5a2},
journal = {J Biol Chem},
keywords = {Animals; Buffers; Calcium Cattle; Channel, Conformation; Cryoelectron Isoforms, Microscopy; Models, Molecular; Protein Receptor Release Ryanodine chemistry/ultrastructure chemistry/ultrastructure;},
month = Mar,
number = 13,
pages = {9485--9491},
pdf = {Shar_2000_9485.pdf},
pmid = {10734096},
timestamp = {2009-06-03T11:21:30.000+0200},
title = {Three-dimensional structure of ryanodine receptor isoform three in
two conformational states as visualized by cryo-electron microscopy.},
url = {http://www.jbc.org/cgi/content/full/275/13/9485},
volume = 275,
year = 2000
}