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TitleCa inactivation of the mammalian ryanodine receptor type 1 in a lipidic environment revealed by cryo-EM.
Journal, issue, pagesElife, Vol. 11, Year 2022
Publish dateMar 8, 2022
AuthorsAshok R Nayak / Montserrat Samsó /
PubMed AbstractActivation of the intracellular Ca channel ryanodine receptor (RyR) triggers a cytosolic Ca surge, while elevated cytosolic Ca inhibits the channel in a negative feedback mechanism. Cryogenic ...Activation of the intracellular Ca channel ryanodine receptor (RyR) triggers a cytosolic Ca surge, while elevated cytosolic Ca inhibits the channel in a negative feedback mechanism. Cryogenic electron microscopy of rabbit RyR1 embedded in nanodiscs under partially inactivating Ca conditions revealed an open and a closed-inactivated conformation. Ca binding to the high-affinity site engages the central and C-terminal domains into a block, which pries the S6 four-helix bundle open. Further rotation of this block pushes S6 toward the central axis, closing (inactivating) the channel. Main characteristics of the Ca-inactivated conformation are downward conformation of the cytoplasmic assembly and tightly knit subunit interface contributed by a fully occupied Ca activation site, two inter-subunit resolved lipids, and two salt bridges between the EF hand domain and the S2-S3 loop validated by disease-causing mutations. The structural insight illustrates the prior Ca activation prerequisite for Ca inactivation and provides for a seamless transition from inactivated to closed conformations.
External linksElife / PubMed:35257661 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 4.3 Å
Structure data

EMDB-22597:
Focused cryo-EM map of rabbit RyR1 central and transmembrane domains in the presence of AMP-PCP in nanodisc
Method: EM (single particle) / Resolution: 3.76 Å

22616

7k0t

EMDB-22616, PDB-7k0t:
Cryo-EM structure of rabbit RyR1 in the presence of AMP-PCP in nanodisc
Method: EM (single particle) / Resolution: 4.3 Å

25828

7tdg

EMDB-25828, PDB-7tdg:
Rabbit RyR1 with AMP-PCP and high Ca2+ embedded in nanodisc in inactivated conformation (Dataset-A)
Method: EM (single particle) / Resolution: 3.8 Å

25829

7tdh

EMDB-25829, PDB-7tdh:
Rabbit RyR1 with AMP-PCP and high Ca2+ embedded in nanodisc in open conformation
Method: EM (single particle) / Resolution: 4.0 Å

25830

7tdi

EMDB-25830, PDB-7tdi:
Rabbit RyR1 with AMP-PCP and high Ca2+ embedded in nanodisc in closed-inactivated conformation class 2 (Dataset-A)
Method: EM (single particle) / Resolution: 3.3 Å

25831

7tdj

EMDB-25831, PDB-7tdj:
Rabbit RyR1 with AMP-PCP and high Ca2+ embedded in nanodisc in closed-inactivated conformation class 1(Dataset-A)
Method: EM (single particle) / Resolution: 3.7 Å

25832

7tdk

EMDB-25832, PDB-7tdk:
Rabbit RyR1 with AMP-PCP and high Ca2+ embedded in nanodisc in closed-inactivated conformation class 3 (Dataset-A)
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-25833:
Rabbit RyR1 with AMP-PCP and high Ca2+ embedded in nanodisc in inactivated conformation (Dataset-B)
Method: EM (single particle) / Resolution: 4.1 Å

Chemicals

ChemComp-ACP:
PHOSPHOMETHYLPHOSPHONIC ACID ADENYLATE ESTER / AMP-PCP, energy-carrying molecule analogue*YM

ChemComp-ZN:
Unknown entry

ChemComp-CA:
Unknown entry

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

ChemComp-HOH:
WATER

Source
  • oryctolagus cuniculus (rabbit)
  • rabbit (rabbit)
KeywordsTRANSPORT PROTEIN / Ryanodine Receptor / RyR1 / Intracellular Calcium channel / Excitation-Contraction coupling / ATP / Ca2+ / Inactivation

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