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TitleResting-State Structure and Gating Mechanism of a Voltage-Gated Sodium Channel.
Journal, issue, pagesCell, Vol. 178, Issue 4, Page 993-1003.e12, Year 2019
Publish dateAug 8, 2019
AuthorsGoragot Wisedchaisri / Lige Tonggu / Eedann McCord / Tamer M Gamal El-Din / Liguo Wang / Ning Zheng / William A Catterall /
PubMed AbstractVoltage-gated sodium (Na) channels initiate action potentials in nerve, muscle, and other electrically excitable cells. The structural basis of voltage gating is uncertain because the resting state ...Voltage-gated sodium (Na) channels initiate action potentials in nerve, muscle, and other electrically excitable cells. The structural basis of voltage gating is uncertain because the resting state exists only at deeply negative membrane potentials. To stabilize the resting conformation, we inserted voltage-shifting mutations and introduced a disulfide crosslink in the VS of the ancestral bacterial sodium channel NaAb. Here, we present a cryo-EM structure of the resting state and a complete voltage-dependent gating mechanism. The S4 segment of the VS is drawn intracellularly, with three gating charges passing through the transmembrane electric field. This movement forms an elbow connecting S4 to the S4-S5 linker, tightens the collar around the S6 activation gate, and prevents its opening. Our structure supports the classical "sliding helix" mechanism of voltage sensing and provides a complete gating mechanism for voltage sensor function, pore opening, and activation-gate closure based on high-resolution structures of a single sodium channel protein.
External linksCell / PubMed:31353218 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.75 - 4.0 Å
Structure data

20265

6p6w

EMDB-20265, PDB-6p6w:
Cryo-EM structure of voltage-gated sodium channel NavAb N49K/L109A/M116V/G94C/Q150C disulfide crosslinked mutant in the resting state
Method: EM (single particle) / Resolution: 4.0 Å

PDB-6p6x:
Crystal structure of voltage-gated sodium channel NavAb G94C/Q150C mutant in the activated state
Method: X-RAY DIFFRACTION / Resolution: 2.75 Å

PDB-6p6y:
Crystal structure of voltage-gated sodium channel NavAb V100C/Q150C disulfide crosslinked mutant in the activated state
Method: X-RAY DIFFRACTION / Resolution: 2.89 Å

Chemicals

ChemComp-PX4:
1,2-DIMYRISTOYL-SN-GLYCERO-3-PHOSPHOCHOLINE / DMPC, phospholipid*YM / Dimyristoylphosphatidylcholine

ChemComp-CPS:
3-[(3-CHOLAMIDOPROPYL)DIMETHYLAMMONIO]-1-PROPANESULFONATE / detergent*YM / CHAPS detergent

Source
  • Arcobacter butzleri RM4018 (bacteria)
  • arcobacter butzleri (strain rm4018) (bacteria)
  • escherichia coli k-12 (bacteria)
Keywordsmembrane protein / metal transport / Ion channel / ion transport protein

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