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- EMDB-22150: CryoEM structure of GIRK2PIP2* - G protein-gated inwardly rectify... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-22150 | |||||||||
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Title | CryoEM structure of GIRK2PIP2* - G protein-gated inwardly rectifying potassium channel GIRK2 with PIP2 | |||||||||
![]() | CryoEM structure of GIRK2PIP2* - G protein-gated inwardly rectifying potassium channel GIRK2 with PIP2 | |||||||||
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![]() | GIRK / inwardly rectifying potassium channel / cholesterol / lipids / PIP2 / TRANSPORT PROTEIN | |||||||||
Function / homology | ![]() G-protein activated inward rectifier potassium channel activity / monoatomic ion channel complex Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.2 Å | |||||||||
![]() | Mathiharan YK / Glaaser IW / Skiniotis G / Slesinger PA | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural insights into GIRK2 channel modulation by cholesterol and PIP. Authors: Yamuna Kalyani Mathiharan / Ian W Glaaser / Yulin Zhao / Michael J Robertson / Georgios Skiniotis / Paul A Slesinger / ![]() Abstract: G-protein-gated inwardly rectifying potassium (GIRK) channels are important for determining neuronal excitability. In addition to G proteins, GIRK channels are potentiated by membrane cholesterol, ...G-protein-gated inwardly rectifying potassium (GIRK) channels are important for determining neuronal excitability. In addition to G proteins, GIRK channels are potentiated by membrane cholesterol, which is elevated in the brains of people with neurodegenerative diseases such as Alzheimer's dementia and Parkinson's disease. The structural mechanism of cholesterol modulation of GIRK channels is not well understood. In this study, we present cryo- electron microscopy (cryoEM) structures of GIRK2 in the presence and absence of the cholesterol analog cholesteryl hemisuccinate (CHS) and phosphatidylinositol 4,5-bisphosphate (PIP). The structures reveal that CHS binds near PIP in lipid-facing hydrophobic pockets of the transmembrane domain. Our structural analysis suggests that CHS stabilizes PIP interaction with the channel and promotes engagement of the cytoplasmic domain onto the transmembrane region. Mutagenesis of one of the CHS binding pockets eliminates cholesterol-dependent potentiation of GIRK2. Elucidating the structural mechanisms underlying cholesterol modulation of GIRK2 channels could facilitate the development of therapeutics for treating neurological diseases. VIDEO ABSTRACT. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 65.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 15.3 KB 15.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.4 KB | Display | ![]() |
Images | ![]() | 136.4 KB | ||
Filedesc metadata | ![]() | 6 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 531.3 KB | Display | ![]() |
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Full document | ![]() | 530.9 KB | Display | |
Data in XML | ![]() | 11 KB | Display | |
Data in CIF | ![]() | 14.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6xeuMC ![]() 6xevC C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | CryoEM structure of GIRK2PIP2* - G protein-gated inwardly rectifying potassium channel GIRK2 with PIP2 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.85 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : G protein-gated inwardly rectifying potassium channel (GIRK2)
Entire | Name: G protein-gated inwardly rectifying potassium channel (GIRK2) |
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Components |
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-Supramolecule #1: G protein-gated inwardly rectifying potassium channel (GIRK2)
Supramolecule | Name: G protein-gated inwardly rectifying potassium channel (GIRK2) type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 Details: The cryoEM structure obtained in the presence of modulator PIP2. |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: G protein-activated inward rectifier potassium channel 2
Macromolecule | Name: G protein-activated inward rectifier potassium channel 2 type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 39.061957 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MAKRKIQRYV RKDGKCNVHH GNVRETYRYL TDIFTTLVDL KWRFNLLIFV MVYTVTWLFF GMIWWLIAYI RGDMDHIEDP SWTPCVTNL NGFVSAFLFS IETETTIGYG YRVITDKCPE GIILLLIQSV LGSIVNAFMV GCMFVKISQP KKRAETLVFS T HAVISMRD ...String: MAKRKIQRYV RKDGKCNVHH GNVRETYRYL TDIFTTLVDL KWRFNLLIFV MVYTVTWLFF GMIWWLIAYI RGDMDHIEDP SWTPCVTNL NGFVSAFLFS IETETTIGYG YRVITDKCPE GIILLLIQSV LGSIVNAFMV GCMFVKISQP KKRAETLVFS T HAVISMRD GKLCLMFRVG DLRNSHIVEA SIRAKLIKSK QTSEGEFIPL NQTDINVGYY TGDDRLFLVS PLIISHEINQ QS PFWEISK AQLPKEELEI VVILEGMVEA TGMTCQARSS YITSEILWGY RFTPVLTLED GFYEVDYNSF HETYETSTPS LSA KELAEL ANRAESNSLE VLFQ UniProtKB: G protein-activated inward rectifier potassium channel 2 |
-Macromolecule #2: [(2R)-2-octanoyloxy-3-[oxidanyl-[(1R,2R,3S,4R,5R,6S)-2,3,6-tris(o...
Macromolecule | Name: [(2R)-2-octanoyloxy-3-[oxidanyl-[(1R,2R,3S,4R,5R,6S)-2,3,6-tris(oxidanyl)-4,5-diphosphonooxy-cyclohexyl]oxy-phosphoryl]oxy-propyl] octanoate type: ligand / ID: 2 / Number of copies: 4 / Formula: PIO |
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Molecular weight | Theoretical: 746.566 Da |
Chemical component information | ![]() ChemComp-PIO: |
-Macromolecule #3: POTASSIUM ION
Macromolecule | Name: POTASSIUM ION / type: ligand / ID: 3 / Number of copies: 8 / Formula: K |
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Molecular weight | Theoretical: 39.098 Da |
-Macromolecule #4: SODIUM ION
Macromolecule | Name: SODIUM ION / type: ligand / ID: 4 / Number of copies: 4 |
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Molecular weight | Theoretical: 22.99 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 7 mg/mL | |||||||||||||||||||||
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Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE | |||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV | |||||||||||||||||||||
Details | GIRK2PIP2* with the modulator PIP2 |
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Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number real images: 6480 / Average exposure time: 3.0 sec. / Average electron dose: 83.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.8 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |