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Structure paper

TitleLipids modulate the open probability of RyR1 under cryo-EM conditions.
Journal, issue, pagesStructure, Vol. 33, Issue 12, Page 2029-22040.e3, Year 2025
Publish dateDec 4, 2025
AuthorsChenyao Li / Rouslan G Efremov /
PubMed AbstractRyanodine receptors (RyRs) are intracellular tetrameric ion channels responsible for Ca release from the sarcoplasmic and endoplasmic reticulum. Ryanodine receptor 1 (RyR1) isoform, critical for ...Ryanodine receptors (RyRs) are intracellular tetrameric ion channels responsible for Ca release from the sarcoplasmic and endoplasmic reticulum. Ryanodine receptor 1 (RyR1) isoform, critical for muscle contraction, has been studied most extensively. While cryoelectron microscopy (cryo-EM) has been instrumental in revealing near-atomic details of RyR gating mechanisms, the open probability of RyR1 under cryo-EM conditions is notably lower than that observed in electrophysiological studies. Here, we present a cryo-EM study examining the open probability of RyR1 solubilized in CHAPS with varying lipid concentrations. We found that increasing lipid concentration from 0.001% to 0.05% raised the RyR1 open probability from 16% to 84%, whereas RyR1 reconstituted into lipid nanodiscs remained closed. We modeled 72 lipid molecules in the map reconstructed at the highest lipid concentration. These findings demonstrate the important role of lipids in modulating the open fraction of solubilized RyR1 channels under cryo-EM conditions and suggest optimal lipid mimetics for structural studies of RyR1 gating.
External linksStructure / PubMed:41027431
MethodsEM (single particle)
Resolution3.3 - 3.6 Å
Structure data

52085

9heo

EMDB-52085, PDB-9heo:
Open-state RyR1 in 0.05% POPC micelles, in complex with a nanobody and FKBP
Method: EM (single particle) / Resolution: 3.4 Å

52091

9heq

EMDB-52091, PDB-9heq:
Open-state RyR1 in 0.01% POPC micelles, in complex with a nanobody and FKBP12
Method: EM (single particle) / Resolution: 3.6 Å

53834

9r8o

EMDB-53834, PDB-9r8o:
Primed-state RyR1 in 0.05% POPC micelles, in complex with a nanobody and FKBP
Method: EM (single particle) / Resolution: 3.3 Å

53924

9rcw

EMDB-53924, PDB-9rcw:
Primed-state RyR1 in 0.01% POPC micelles, in complex with a nanobody and FKBP12
Method: EM (single particle) / Resolution: 3.3 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

ChemComp-CFF:
CAFFEINE / medication*YM

ChemComp-CA:
Unknown entry

ChemComp-POV:
(2S)-3-(hexadecanoyloxy)-2-[(9Z)-octadec-9-enoyloxy]propyl 2-(trimethylammonio)ethyl phosphate / phospholipid*YM

Source
  • oryctolagus cuniculus (rabbit)
  • Vicugna pacos (alpaca)
  • lama glama (llama)
KeywordsTRANSPORT PROTEIN / Ion channel / Ca2+ / tetramer

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