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PMID: 22913516 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Structural determination of the phosphorylation domain of the ryanodine receptor.

The FEBS journal ·Vol. 279 ·No. 20 ·2012-10-00 ·Pages 3952-64

Sharma P, Ishiyama N, Nair U, Li W, Dong A, Miyake T, Wilson A, Ryan T, MacLennan DH, Kislinger T, Ikura M, Dhe-Paganon S, Gramolini AO

Abstract

The ryanodine receptor (RyR) is a large, homotetrameric sarcoplasmic reticulum membrane protein that is essential for Ca(2+) cycling in both skeletal and cardiac muscle. Genetic mutations in RyR1 are associated with severe conditions including malignant hyperthermia (MH) and central core disease. One phosphorylation site (Ser 2843) has been identified in a segment of RyR1 flanked by two RyR motifs, which are found exclusively in all RyR isoforms as closely associated tandem (or paired) motifs, and are named after the protein itself. These motifs also contain six known MH mutations. In this study, we designed, expressed and purified the tandem RyR motifs, and show that this domain contains a putative binding site for the Ca(2+)/calmodulin-dependent protein kinase β isoform. We present a 2.2 Å resolution crystal structure of the RyR domain revealing a two-fold, symmetric, extended four-helix bundle stabilized by a β sheet. Using mathematical modelling, we fit our crystal structure within a tetrameric electron microscopy (EM) structure of native RyR1, and propose that this domain is localized in the RyR clamp region, which is absent in its cousin protein inositol 1,4,5-trisphosphate receptor.

MeSH Terms
Amino Acid Motifs/genetics Amino Acid Sequence Animals Binding Sites Blotting, Western Calcium-Calmodulin-Dependent Protein Kinase Type 2/chemistry,metabolism Cryoelectron Microscopy Crystallography, X-Ray HEK293 Cells Humans Models, Molecular Molecular Sequence Data Mutation Phosphorylation Protein Binding Protein Isoforms/chemistry,genetics,metabolism Protein Structure, Secondary Protein Structure, Tertiary Rabbits Ryanodine Receptor Calcium Release Channel/chemistry,genetics,metabolism Sequence Homology, Amino Acid
Chemicals
Protein Isoforms Ryanodine Receptor Calcium Release Channel Calcium-Calmodulin-Dependent Protein Kinase Type 2
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Sharma Parveen
Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
Ishiyama Noboru
Nair Usha
Li Wenping
Dong Aiping
Miyake Tetsuaki
Wilson Aaron
Ryan Tim
MacLennan David H
Kislinger Thomas
Ikura Mitsuhiko
Dhe-Paganon Sirano
Gramolini Anthony O
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Article Info
Journal
The FEBS journal
Abbr.
FEBS J
ISSN
1742-4658
DOI
10.1111/j.1742-4658.2012.08755.x
Published
2012-10-00
Epub
2012-00-11
Pages
3952-64
Language
English
Region
England
NLM ID
101229646
PMCID
PMC3712973
Subset
IM
Grants
Wellcome Trust · United Kingdom
Canadian Institutes of Health Research · 84267-1 · Canada
CIHR · MOP-3399 · Canada
CIHR · MOP-84267 · Canada
Databases
PDB
3RQR
External Links
PubMed source DOI: 10.1111/j.1742-4658.2012.08755.x PMC Full Text View journal details
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