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- EMDB-16819: Chaetomium thermophilum Get1/Get2 heterotetramer in complex with ... -
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Open data
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Basic information
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Title | Chaetomium thermophilum Get1/Get2 heterotetramer in complex with a Get3 dimer (nanodisc) | |||||||||
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![]() | membrane protein insertion / GET pathway / tail anchored membrane protein / MEMBRANE PROTEIN | |||||||||
Function / homology | ![]() GET complex / tail-anchored membrane protein insertion into ER membrane / protein insertion into ER membrane / post-translational protein targeting to endoplasmic reticulum membrane / retrograde vesicle-mediated transport, Golgi to endoplasmic reticulum / protein-membrane adaptor activity / endoplasmic reticulum membrane / ATP hydrolysis activity / ATP binding / membrane / metal ion binding Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.7 Å | |||||||||
![]() | McDowell MA / Wild K / Sinning I | |||||||||
Funding support | ![]()
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![]() | ![]() Title: The GET insertase exhibits conformational plasticity and induces membrane thinning. Authors: Melanie A McDowell / Michael Heimes / Giray Enkavi / Ákos Farkas / Daniel Saar / Klemens Wild / Blanche Schwappach / Ilpo Vattulainen / Irmgard Sinning / ![]() ![]() Abstract: The eukaryotic guided entry of tail-anchored proteins (GET) pathway mediates the biogenesis of tail-anchored (TA) membrane proteins at the endoplasmic reticulum. In the cytosol, the Get3 chaperone ...The eukaryotic guided entry of tail-anchored proteins (GET) pathway mediates the biogenesis of tail-anchored (TA) membrane proteins at the endoplasmic reticulum. In the cytosol, the Get3 chaperone captures the TA protein substrate and delivers it to the Get1/Get2 membrane protein complex (GET insertase), which then inserts the substrate via a membrane-embedded hydrophilic groove. Here, we present structures, atomistic simulations and functional data of human and Chaetomium thermophilum Get1/Get2/Get3. The core fold of the GET insertase is conserved throughout eukaryotes, whilst thinning of the lipid bilayer occurs in the vicinity of the hydrophilic groove to presumably lower the energetic barrier of membrane insertion. We show that the gating interaction between Get2 helix α3' and Get3 drives conformational changes in both Get3 and the Get1/Get2 membrane heterotetramer. Thus, we provide a framework to understand the conformational plasticity of the GET insertase and how it remodels its membrane environment to promote substrate insertion. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 59.6 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 22.1 KB 22.1 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 8.9 KB | Display | ![]() |
Images | ![]() | 53.8 KB | ||
Masks | ![]() | 64 MB | ![]() | |
Filedesc metadata | ![]() | 6.7 KB | ||
Others | ![]() ![]() ![]() | 59.6 MB 59.3 MB 59.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8odvMC ![]() 8cqzC ![]() 8cr1C ![]() 8cr2C ![]() 8oduC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 1.11 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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-Additional map: Final map low-pass filtered to 6.5 A
File | emd_16819_additional_1.map | ||||||||||||
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Annotation | Final map low-pass filtered to 6.5 A | ||||||||||||
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-Half map: #2
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Density Histograms |
-Half map: #1
File | emd_16819_half_map_2.map | ||||||||||||
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Sample components
-Entire : Chaetomium thermophilum Get1/Get2 heterotetramer in complex with ...
Entire | Name: Chaetomium thermophilum Get1/Get2 heterotetramer in complex with a Get3 dimer |
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Components |
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-Supramolecule #1: Chaetomium thermophilum Get1/Get2 heterotetramer in complex with ...
Supramolecule | Name: Chaetomium thermophilum Get1/Get2 heterotetramer in complex with a Get3 dimer type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 Details: Get2-Get1 was expressed as a fusion protein in S. cerevisiae and Get3 was expressed in E. coli. The complex components were purified and reconstituted in vitro. |
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Molecular weight | Theoretical: 165 KDa |
-Supramolecule #2: Tail-anchored protein insertase Get1 and Get2
Supramolecule | Name: Tail-anchored protein insertase Get1 and Get2 / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #2 |
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Source (natural) | Organism: ![]() |
-Supramolecule #3: Dimeric ATPase Get3
Supramolecule | Name: Dimeric ATPase Get3 / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: ATPase GET3
Macromolecule | Name: ATPase GET3 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO / EC number: Hydrolases; Acting on acid anhydrides |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 37.127391 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MEPTLQSILD QRSLRWIFVG GKGGVGKTTT SCSLAIQLAK VRRSVLLLST DPAHNLSDAF SQKFGKEARL VEGFDNLYAM EIDPNGSMQ DLLAGQTGDG DAGMGGVGVM QDLAYAIPGI DEAMSFAEVL KQVNSLSYET IVFDTAPTGH TLRFLQFPTV L EKALAKVS ...String: MEPTLQSILD QRSLRWIFVG GKGGVGKTTT SCSLAIQLAK VRRSVLLLST DPAHNLSDAF SQKFGKEARL VEGFDNLYAM EIDPNGSMQ DLLAGQTGDG DAGMGGVGVM QDLAYAIPGI DEAMSFAEVL KQVNSLSYET IVFDTAPTGH TLRFLQFPTV L EKALAKVS QLSGQYGSLL NGILGGSGTL PNGQTLSDVM EKLDSLRVTI SEVNAQFKDE RLTTFVCVCI PEFLSLYETE RM IQELANY GIDTHCIVVN QLLFPKPGSD CEQCTARRRM QKKYLDQIEE LYDEEFNVVK MPLLVEEVRG KERLEKFSEM LIK PFVPPE WSHPQFEK UniProtKB: Arsenite translocating ATPase-like protein |
-Macromolecule #2: Protein GET2,Protein GET1
Macromolecule | Name: Protein GET2,Protein GET1 / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 45.573727 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MGRPTPLWRF LHTLLAVALG LAVIMLSPFG GTKLERDRAA AAVAGSASER EWLASLTDSY PLVKTGLGGG LFWAFATGEA ILLGTRWLF LSKKKKAATA AAKVNNNNGE GDDAELDSVE QAIELALEFF PAIRQPVEYL RPKVAVAMRY VDVGMTLWRD V MLALFVLG ...String: MGRPTPLWRF LHTLLAVALG LAVIMLSPFG GTKLERDRAA AAVAGSASER EWLASLTDSY PLVKTGLGGG LFWAFATGEA ILLGTRWLF LSKKKKAATA AAKVNNNNGE GDDAELDSVE QAIELALEFF PAIRQPVEYL RPKVAVAMRY VDVGMTLWRD V MLALFVLG AVAWWRAGSG SENLYFQSGS GSMSLLLVIF LLELVVQLVN TIGAKTINNL LWRFYLSIPG SPLAKDFAEQ RA KQKEYLQ VRHDLNATSS QDEFAKWARL QRKHDKLMDE LEKKKSQLDA HRTSFSRKLT IYRWILTRGM QWFLCFWFSS QPM FWLPYG WFPYWVEWLV SFPNAPMGSV SIVVWQSACS GVLALVIEAV MAVVRYTGGT GMQKQRQPVP AAGGAPGTSK KDLG SGSLE VLFQ UniProtKB: Uncharacterized protein, Protein GET1 |
-Macromolecule #3: ZINC ION
Macromolecule | Name: ZINC ION / type: ligand / ID: 3 / Number of copies: 1 / Formula: ZN |
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Molecular weight | Theoretical: 65.409 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 | 2.8 mg/mL | ||||||
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Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil R2/1 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE | ||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 279 K / Instrument: FEI VITROBOT MARK IV | ||||||
Details | Complex stabilised in a Msp1E3D1 bound nanodisc |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 11997 / Average exposure time: 2.6 sec. / Average electron dose: 60.8 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: 2.4 µm / Nominal defocus min: 1.2 µm |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: RIGID BODY FIT |
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Output model | ![]() PDB-8odv: |