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Basic information
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Title | Structure of AHIR determined by cryoEM at 100 keV | |||||||||||||||||||||
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![]() | OXIDOREDUCTASE | |||||||||||||||||||||
Function / homology | ![]() ketol-acid reductoisomerase (NADP+) / ketol-acid reductoisomerase activity / valine biosynthetic process / isoleucine biosynthetic process / NADP binding / magnesium ion binding Similarity search - Function | |||||||||||||||||||||
Biological species | ![]() ![]() | |||||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||||||||||||||
![]() | McMullan G / Naydenova K / Mihaylov D / Peet MJ / Wilson H / Yamashita K / Dickerson JL / Chen S / Cannone G / Lee Y ...McMullan G / Naydenova K / Mihaylov D / Peet MJ / Wilson H / Yamashita K / Dickerson JL / Chen S / Cannone G / Lee Y / Hutchings KA / Gittins O / Sobhy M / Wells T / El-Gomati MM / Dalby J / Meffert M / Schulze-Briese C / Henderson R / Russo CJ | |||||||||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structure determination by cryoEM at 100 keV. Authors: Greg McMullan / Katerina Naydenova / Daniel Mihaylov / Keitaro Yamashita / Mathew J Peet / Hugh Wilson / Joshua L Dickerson / Shaoxia Chen / Giuseppe Cannone / Yang Lee / Katherine A ...Authors: Greg McMullan / Katerina Naydenova / Daniel Mihaylov / Keitaro Yamashita / Mathew J Peet / Hugh Wilson / Joshua L Dickerson / Shaoxia Chen / Giuseppe Cannone / Yang Lee / Katherine A Hutchings / Olivia Gittins / Mohamed A Sobhy / Torquil Wells / Mohamed M El-Gomati / Jason Dalby / Matthias Meffert / Clemens Schulze-Briese / Richard Henderson / Christopher J Russo / ![]() ![]() ![]() Abstract: Electron cryomicroscopy can, in principle, determine the structures of most biological molecules but is currently limited by access, specimen preparation difficulties, and cost. We describe a purpose- ...Electron cryomicroscopy can, in principle, determine the structures of most biological molecules but is currently limited by access, specimen preparation difficulties, and cost. We describe a purpose-built instrument operating at 100 keV-including advances in electron optics, detection, and processing-that makes structure determination fast and simple at a fraction of current costs. The instrument attains its theoretical performance limits, allowing atomic resolution imaging of gold test specimens and biological molecular structure determination in hours. We demonstrate its capabilities by determining the structures of eleven different specimens, ranging in size from 140 kDa to 2 MDa, using a fraction of the data normally required. CryoEM with a microscope designed specifically for high-efficiency, on-the-spot imaging of biological molecules will expand structural biology to a wide range of previously intractable problems. | |||||||||||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 19.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 17.3 KB 17.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 14.9 KB | Display | ![]() |
Images | ![]() | 97.8 KB | ||
Masks | ![]() | 282.6 MB | ![]() | |
Filedesc metadata | ![]() | 6 KB | ||
Others | ![]() ![]() | 225.4 MB 225.2 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 806.2 KB | Display | ![]() |
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Full document | ![]() | 805.7 KB | Display | |
Data in XML | ![]() | 22.4 KB | Display | |
Data in CIF | ![]() | 29.4 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8pveMC ![]() 8pv9C ![]() 8pvaC ![]() 8pvbC ![]() 8pvcC ![]() 8pvdC ![]() 8pvfC ![]() 8pvgC ![]() 8pvhC ![]() 8pviC ![]() 8pvjC 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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Related items in Molecule of the Month |
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Map
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Voxel size | X=Y=Z: 0.8315 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 : Acetohydroxy acid isomeroreductase
Entire | Name: Acetohydroxy acid isomeroreductase |
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Components |
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-Supramolecule #1: Acetohydroxy acid isomeroreductase
Supramolecule | Name: Acetohydroxy acid isomeroreductase / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: Ketol-acid reductoisomerase (NADP(+))
Macromolecule | Name: Ketol-acid reductoisomerase (NADP(+)) / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: ketol-acid reductoisomerase (NADP+) |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 37.994246 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MTGTMKIYYE HDADLGFILG KKVAVLGFGS QGHAHAHNLK DSGVDVRVGL RKGSRSWEKA EAAGLRVLPV AEAVREADVV MVLLPDEKQ AQVYREEVEP NLKEGGALAF AHGFNVHFGQ IKPRKDLDVW MVAPKGPGHL VRSEYGGGSN WSHPQFEKRP P GGSGVPAL ...String: MTGTMKIYYE HDADLGFILG KKVAVLGFGS QGHAHAHNLK DSGVDVRVGL RKGSRSWEKA EAAGLRVLPV AEAVREADVV MVLLPDEKQ AQVYREEVEP NLKEGGALAF AHGFNVHFGQ IKPRKDLDVW MVAPKGPGHL VRSEYGGGSN WSHPQFEKRP P GGSGVPAL VAVHQDASGS AFPTALAYAK AIGAARAGVI ATTFKDETET DLFGEQAVLC GGLTRLIRAG FETLVEAGYP PE MAYFETV HEVKLIVDLI YEAGLKGMRY SISNTAEYGD YTRGDLAVPL EETKRRMREI LRQIQSGEFA REWMLENQVG SPV LEANRK RWAAHPIEEV GSRLRAMMRS UniProtKB: Ketol-acid reductoisomerase (NADP(+)) |
-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: 7.4 |
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 200 |
Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | JEOL 1400/HR + YPS FEG |
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Image recording | Film or detector model: DECTRIS SINGLA (1k x 1k) / Average electron dose: 80.0 e/Å2 |
Electron beam | Acceleration voltage: 100 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm |
Sample stage | Specimen holder model: GATAN 626 SINGLE TILT LIQUID NITROGEN CRYO TRANSFER HOLDER Cooling holder cryogen: NITROGEN |
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Image processing
-Atomic model buiding 1
Refinement | Space: RECIPROCAL |
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Output model | ![]() PDB-8pve: |