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Open data
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Basic information
Entry | ![]() | |||||||||
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Title | Nucleotide-free Get3 in the closed form | |||||||||
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![]() | Tail-anchored membrane protein targeting Deviant Walker A ATPase targeting factor / CHAPERONE | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 8.46 Å | |||||||||
![]() | Fry MY / Clemons Jr WM | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structurally derived universal mechanism for the catalytic cycle of the tail-anchored targeting factor Get3. Authors: Michelle Y Fry / Vladimíra Najdrová / Ailiena O Maggiolo / Shyam M Saladi / Pavel Doležal / William M Clemons / ![]() ![]() Abstract: Tail-anchored (TA) membrane proteins, accounting for roughly 2% of proteomes, are primarily targeted posttranslationally to the endoplasmic reticulum membrane by the guided entry of TA proteins (GET) ...Tail-anchored (TA) membrane proteins, accounting for roughly 2% of proteomes, are primarily targeted posttranslationally to the endoplasmic reticulum membrane by the guided entry of TA proteins (GET) pathway. For this complicated process, it remains unknown how the central targeting factor Get3 uses nucleotide to facilitate large conformational changes to recognize then bind clients while also preventing exposure of hydrophobic surfaces. Here, we identify the GET pathway in Giardia intestinalis and present the structure of the Get3-client complex in the critical postnucleotide-hydrolysis state, demonstrating that Get3 reorganizes the client-binding domain (CBD) to accommodate and shield the client transmembrane helix. Four additional structures of GiGet3, spanning the nucleotide-free (apo) open to closed transition and the ATP-bound state, reveal the details of nucleotide stabilization and occluded CBD. This work resolves key conundrums and allows for a complete model of the dramatic conformational landscape of Get3. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 25.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 14.4 KB 14.4 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 7 KB | Display | ![]() |
Images | ![]() | 48.8 KB | ||
Masks | ![]() | 27 MB | ![]() | |
Filedesc metadata | ![]() | 4.9 KB | ||
Others | ![]() ![]() | 20.7 MB 20.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 919.1 KB | Display | ![]() |
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Full document | ![]() | 918.7 KB | Display | |
Data in XML | ![]() | 12.3 KB | Display | |
Data in CIF | ![]() | 17 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 1.059 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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Density Histograms |
-Half map: #1
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Projections & Slices |
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Density Histograms |
-Half map: #2
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Density Histograms |
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Sample components
-Entire : Nucleotide-free Get3 in a closed conformation
Entire | Name: Nucleotide-free Get3 in a closed conformation |
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Components |
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-Supramolecule #1: Nucleotide-free Get3 in a closed conformation
Supramolecule | Name: Nucleotide-free Get3 in a closed conformation / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() Strain: ATCC 50803 / WB clone C6 |
Molecular weight | Theoretical: 78 KDa |
-Macromolecule #1: ATPase GET3
Macromolecule | Name: ATPase GET3 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MLPSLHDILD QHTYKWIFFG GKGGVGKTTT SSSFSVLMAE TRPNEKFLLL STDPAHNISD AFDQKFGKAP TQVSGIPNLY AMEVDASNEM KSAVEAVQKE TGSAADNDAE SKSEGDMFGG LNDLITCASS FIKDGTFPGM DEMWSFINLI KLIDTNEYST VIFDTAPTGH ...String: MLPSLHDILD QHTYKWIFFG GKGGVGKTTT SSSFSVLMAE TRPNEKFLLL STDPAHNISD AFDQKFGKAP TQVSGIPNLY AMEVDASNEM KSAVEAVQKE TGSAADNDAE SKSEGDMFGG LNDLITCASS FIKDGTFPGM DEMWSFINLI KLIDTNEYST VIFDTAPTGH TLRFLELPET VNKVLEIFTR LKDNMGGMLS MVMQTMGLSQ NDIFGLIDKT YPKIDVVKRI SAEFRDPSLC TFVGVCIPEF LSLYETERLV QRLAVLDMDC HAIVINFVLD ANAATPCSMC RSRARMQNKY IDQINELYDD FNIVLSPLRH DEVRGIANLR DYAETLIKPY RFCWSANPDP SSAK |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.55 mg/mL |
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Buffer | pH: 7.5 |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 9300 / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: DARK FIELD / Nominal magnification: 88000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |