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Conformational dynamics inside amino-terminal disease hotspot of ryanodine receptor
- Source :
- Structure (London, England : 1993). 21(11)
- Publication Year :
- 2013
-
Abstract
- Summary The N-terminal region of both skeletal and cardiac ryanodine receptor is a disease mutation hotspot. Recently, a crystal structure of the RyR1 fragment (residues 1–559) was solved. This N-terminal structure contains three separate domains, A, B, and C, and was docked into a central vestibule in a full-length RyR1 cryo-EM map. Here, we reconstructed three-dimensional cryo-EM structures of two GFP-tagged RyR2s with GFP inserted after residue Glu-310 and Ser-437, respectively. The structures of RyR2 E310-GFP and RyR2 S437-GFP displayed an extra mass on domain B and C, directly validating the predicted docking model. Next, we revealed domain movements in molecular dynamics flexible fitting models in both the closed and open state cryo-EM maps. To further probe the conformational changes, we generated FRET pairs by inserting CFP or YFP in two selected domains, FRET studies of three dual-insertion pairs and three co-expressed single-insertion pairs showed the dynamic structural changes within the N-terminal domains.
- Subjects :
- Recombinant Fusion Proteins
Green Fluorescent Proteins
Plasma protein binding
Biology
Molecular Docking Simulation
Ryanodine receptor 2
Protein Structure, Secondary
Article
03 medical and health sciences
Molecular dynamics
Mice
0302 clinical medicine
Bacterial Proteins
Structural Biology
Caffeine
Fluorescence Resonance Energy Transfer
Animals
Humans
Protein Interaction Domains and Motifs
Molecular Biology
030304 developmental biology
RYR1
0303 health sciences
Ryanodine receptor
Cryoelectron Microscopy
Ryanodine Receptor Calcium Release Channel
Crystallography
Luminescent Proteins
Förster resonance energy transfer
HEK293 Cells
Docking (molecular)
Biophysics
030217 neurology & neurosurgery
Protein Binding
Subjects
Details
- ISSN :
- 18784186
- Volume :
- 21
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- Structure (London, England : 1993)
- Accession number :
- edsair.doi.dedup.....2bc1c7bcf64393c7a6b8986563421212