|Entry||Database: EMDB / ID: 8632|
|Title||CryoEM Structure of a Prokaryotic Cyclic Nucleotide-Gated Ion Channel|
|Map data||Prokaryotic Cyclic Nucleotide-Gated Ion Channel|
|Function/homology||Potassium channel, voltage-dependent, EAG/ELK/ERG / Cyclic nucleotide-binding domain signature 2. / Cyclic nucleotide-binding, conserved site / voltage-gated potassium channel activity / cAMP/cGMP binding motif profile. / Cyclic nucleotide-binding domain / Cyclic nucleotide-binding-like / Ion transport domain / Cyclic nucleotide-binding domain / Ion transport protein ...Potassium channel, voltage-dependent, EAG/ELK/ERG / Cyclic nucleotide-binding domain signature 2. / Cyclic nucleotide-binding, conserved site / voltage-gated potassium channel activity / cAMP/cGMP binding motif profile. / Cyclic nucleotide-binding domain / Cyclic nucleotide-binding-like / Ion transport domain / Cyclic nucleotide-binding domain / Ion transport protein / RmlC-like jelly roll fold / integral component of membrane / Transporter, cation channel family / cyclic nucleotide-binding domain multi-domain protein|
Function and homology information
|Source||Leptospira licerasiae serovar varillal str. var 010 / / bacteria|
|Method||Cryo EM / single particle reconstruction / 4.2 Å resolution|
|Authors||James ZM / Borst AJ|
|Citation||Journal: Proc. Natl. Acad. Sci. U.S.A. / Year: 2017|
Title: CryoEM structure of a prokaryotic cyclic nucleotide-gated ion channel.
Authors: Zachary M James / Andrew J Borst / Yoni Haitin / Brandon Frenz / Frank DiMaio / William N Zagotta / David Veesler
Abstract: Cyclic nucleotide-gated (CNG) and hyperpolarization-activated cyclic nucleotide-regulated (HCN) ion channels play crucial physiological roles in phototransduction, olfaction, and cardiac pace making. ...Cyclic nucleotide-gated (CNG) and hyperpolarization-activated cyclic nucleotide-regulated (HCN) ion channels play crucial physiological roles in phototransduction, olfaction, and cardiac pace making. These channels are characterized by the presence of a carboxyl-terminal cyclic nucleotide-binding domain (CNBD) that connects to the channel pore via a C-linker domain. Although cyclic nucleotide binding has been shown to promote CNG and HCN channel opening, the precise mechanism underlying gating remains poorly understood. Here we used cryoEM to determine the structure of the intact LliK CNG channel isolated from -which shares sequence similarity to eukaryotic CNG and HCN channels-in the presence of a saturating concentration of cAMP. A short S4-S5 linker connects nearby voltage-sensing and pore domains to produce a non-domain-swapped transmembrane architecture, which appears to be a hallmark of this channel family. We also observe major conformational changes of the LliK C-linkers and CNBDs relative to the crystal structures of isolated C-linker/CNBD fragments and the cryoEM structures of related CNG, HCN, and KCNH channels. The conformation of our LliK structure may represent a functional state of this channel family not captured in previous studies.
|Validation Report||PDB-ID: 5v4s|
SummaryFull reportAbout validation report
|Date||Deposition: Mar 10, 2017 / Header (metadata) release: Apr 5, 2017 / Map release: Apr 12, 2017 / Last update: Sep 27, 2017|
Downloads & links
|File||emd_8632.map.gz (map file in CCP4 format, 49949 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 1.3655 Å|
CCP4 map header:
|Entire||Name: LliK / Number of components: 2|
|Mass||Theoretical: 200 kDa|
-Component #1: protein, LliK
|Protein||Name: LliK / Recombinant expression: No|
|Mass||Theoretical: 200 kDa|
|Source||Species: Leptospira licerasiae serovar varillal str. var 010 / / bacteria|
|Source (engineered)||Expression System: Escherichia coli / / bacteria /|
-Component #2: protein, Transporter, cation channel family / cyclic nucleotide-b...
|Protein||Name: Transporter, cation channel family / cyclic nucleotide-binding domain multi-domain protein|
Recombinant expression: No
|Mass||Theoretical: 53.464785 kDa|
|Source (engineered)||Expression System: Leptospira licerasiae serovar varillal str. var 010 / / bacteria|
|Specimen||Specimen state: particle / Method: Cryo EM|
|Sample solution||Specimen conc.: 0.9 mg/ml / pH: 7.9|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 1.4 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 (4k x 4k)|
|Processing||Method: single particle reconstruction / Applied symmetry: C4 (4 fold cyclic) / Number of projections: 18737|
|3D reconstruction||Software: RELION / Resolution: 4.2 Å / Resolution method: FSC 0.143 CUT-OFF|
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