Back to Search Start Over

Conformational dynamics inside amino-terminal disease hotspot of ryanodine receptor

Authors :
Li Zhu
Filip Van Petegem
Xiaowei Zhong
Ying Liu
Terence Wagenknecht
S.R. Wayne Chen
Ruiwu Wang
Zheng Liu
Xing Meng
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.

Details

ISSN :
18784186
Volume :
21
Issue :
11
Database :
OpenAIRE
Journal :
Structure (London, England : 1993)
Accession number :
edsair.doi.dedup.....2bc1c7bcf64393c7a6b8986563421212