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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-22176 | |||||||||
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Title | Full-length human mitochondrial Hsp90 (TRAP1) with ADP-BeF3 | |||||||||
![]() | Hsp90 (TRAP1) NTD-Middle domain dimer with ADP-BeF3 | |||||||||
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Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.42 Å | |||||||||
![]() | Liu YX / Wang F / Agard DA | |||||||||
Funding support | ![]()
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![]() | ![]() Title: General and robust covalently linked graphene oxide affinity grids for high-resolution cryo-EM. Authors: Feng Wang / Yanxin Liu / Zanlin Yu / Sam Li / Shengjie Feng / Yifan Cheng / David A Agard / ![]() Abstract: Affinity grids have great potential to facilitate rapid preparation of even quite impure samples in single-particle cryo-electron microscopy (EM). Yet despite the promising advances of affinity grids ...Affinity grids have great potential to facilitate rapid preparation of even quite impure samples in single-particle cryo-electron microscopy (EM). Yet despite the promising advances of affinity grids over the past decades, no single strategy has demonstrated general utility. Here we chemically functionalize cryo-EM grids coated with mostly one or two layers of graphene oxide to facilitate affinity capture. The protein of interest is tagged using a system that rapidly forms a highly specific covalent bond to its cognate catcher linked to the grid via a polyethylene glycol (PEG) spacer. Importantly, the spacer keeps particles away from both the air-water interface and the graphene oxide surface, protecting them from potential denaturation and rendering them sufficiently flexible to avoid preferential sample orientation concerns. Furthermore, the PEG spacer successfully reduces nonspecific binding, enabling high-resolution reconstructions from a much cruder lysate sample. | |||||||||
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 | ![]() | 59.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 9.9 KB 9.9 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.1 KB | Display | ![]() |
Images | ![]() | 60 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 78.5 KB | Display | ![]() |
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Full document | ![]() | 77.6 KB | Display | |
Data in XML | ![]() | 495 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Hsp90 (TRAP1) NTD-Middle domain dimer with ADP-BeF3 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.14 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 : Trap1 dimer with ADP-BeF3
Entire | Name: Trap1 dimer with ADP-BeF3 |
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Components |
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-Supramolecule #1: Trap1 dimer with ADP-BeF3
Supramolecule | Name: Trap1 dimer with ADP-BeF3 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Molecular weight | Experimental: 147 KDa |
-Macromolecule #1: Human mitochondrial Hsp90 (TRAP1)
Macromolecule | Name: Human mitochondrial Hsp90 (TRAP1) / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: GIDPFTSTQT AEDKEEPLHS IISSTESVQG STSKHEFQAE TKKLLDIVAR SLYSEKEVFI RELISNASDA LEKLRHKLVS DGQALPEMEI HLQTNAEKGT ITIQDTGIGM TQEELVSNLG TIARSGSKAF LDALQNQAEA SSKIIGQFGV GFYSAFMVAD RVEVYSRSAA ...String: GIDPFTSTQT AEDKEEPLHS IISSTESVQG STSKHEFQAE TKKLLDIVAR SLYSEKEVFI RELISNASDA LEKLRHKLVS DGQALPEMEI HLQTNAEKGT ITIQDTGIGM TQEELVSNLG TIARSGSKAF LDALQNQAEA SSKIIGQFGV GFYSAFMVAD RVEVYSRSAA PGSLGYQWLS DGSGVFEIAE ASGVRTGTKI IIHLKSDCKE FSSEARVRDV VTKYSNFVSF PLYLNGRRMN TLQAIWMMDP KDVGEWQHEE FYRYVAQAHD KPRYTLHYKT DAPLNIRSIF YVPDMKPSMF DVSRELGSSV ALYSRKVLIQ TKATDILPKW LRFIRGVVDS EDIPLNLSRE LLQESALIRK LRDVLQQRLI KFFIDQSKKD AEKYAKFFED YGLFMREGIV TATEQEVKED IAKLLRYESS ALPSGQLTSL SEYASRMRAG TRNIYYLCAP NRHLAEHSPY YEAMKKKDTE VLFCFEQFDE LTLLHLREFD KKKLISVETD IVVDHYKEEK FEDRSPAAEC LSEKETEELM AWMRNVLGSR VTNVKVTLRL DTHPAMVTVL EMGAARHFLR MQQLAKTQEE RAQLLQPTLE INPRHALIKK LNQLRASEPG LAQLLVDQIY ENAMIAAGLV DDPRAMVGRL NELLVKALER HGGSGSGSSA MVDTLSGLSS EQGQSGDMTI EEDSATHIKF SKRDEDGKEL AGATMELRDS SGKTISTWIS DGQVKDFYLY PGKYTFVETA APDGYEVATA ITFTVNEQGQ VTVNGKATKG DAHI |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 74.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Talos Arctica / 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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