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- EMDB-26762: Locally refined cryoEM map of Azotobacter vinelandii FeP in a 1:1... -
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Basic information
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Title | Locally refined cryoEM map of Azotobacter vinelandii FeP in a 1:1 complex (ADP/ATP-bound) with MoFeP during catalytic N2 reduction | |||||||||||||||
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Biological species | ![]() ![]() | |||||||||||||||
Method | ![]() ![]() | |||||||||||||||
![]() | Rutledge HL / Cook B / Tezcan FA / Herzik MA | |||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structures of the nitrogenase complex prepared under catalytic turnover conditions. Authors: Hannah L Rutledge / Brian D Cook / Hoang P M Nguyen / Mark A Herzik / F Akif Tezcan / ![]() Abstract: The enzyme nitrogenase couples adenosine triphosphate (ATP) hydrolysis to the multielectron reduction of atmospheric dinitrogen into ammonia. Despite extensive research, the mechanistic details of ...The enzyme nitrogenase couples adenosine triphosphate (ATP) hydrolysis to the multielectron reduction of atmospheric dinitrogen into ammonia. Despite extensive research, the mechanistic details of ATP-dependent energy transduction and dinitrogen reduction by nitrogenase are not well understood, requiring new strategies to monitor its structural dynamics during catalytic action. Here, we report cryo-electron microscopy structures of the nitrogenase complex prepared under enzymatic turnover conditions. We observe that asymmetry governs all aspects of the nitrogenase mechanism, including ATP hydrolysis, protein-protein interactions, and catalysis. Conformational changes near the catalytic iron-molybdenum cofactor are correlated with the nucleotide-hydrolysis state of the enzyme. | |||||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 180.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 21 KB 21 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 12.7 KB | Display | ![]() |
Images | ![]() | 59.8 KB | ||
Masks | ![]() | 216 MB | ![]() | |
Filedesc metadata | ![]() | 5.5 KB | ||
Others | ![]() ![]() ![]() | 107.9 MB 200.7 MB 200.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | deepEMhanced sharpened map | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.835 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: unsharpened map
File | emd_26762_additional_1.map | ||||||||||||
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Annotation | unsharpened map | ||||||||||||
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-Half map: gold-standard half map A
File | emd_26762_half_map_1.map | ||||||||||||
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Annotation | gold-standard half map A | ||||||||||||
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-Half map: gold-standard half map B
File | emd_26762_half_map_2.map | ||||||||||||
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Annotation | gold-standard half map B | ||||||||||||
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Sample components
-Entire : Locally refined cryoEM map of Azotobacter vinelandii FeP in a 1:1...
Entire | Name: Locally refined cryoEM map of Azotobacter vinelandii FeP in a 1:1 complex (ADP/ATP-bound) with MoFeP during catalytic N2 reduction |
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Components |
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-Supramolecule #1: Locally refined cryoEM map of Azotobacter vinelandii FeP in a 1:1...
Supramolecule | Name: Locally refined cryoEM map of Azotobacter vinelandii FeP in a 1:1 complex (ADP/ATP-bound) with MoFeP during catalytic N2 reduction type: complex / ID: 1 / Parent: 0 / Macromolecule list: all Details: Wild-type MoFeP and FeP were purified from the native organism, Azotobacter vinelandii. This map is the locally refined map for the FeP portion of the 1:1 complex. |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 63 KDa |
-Macromolecule #1: Nitrogenase iron protein gamma chain
Macromolecule | Name: Nitrogenase iron protein gamma chain / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: MAMRQCAIYG KGGIGKSTTT QNLVAALAEM GKKVMIVGCD PKADSTRLIL HSKAQNTIME MAAEAGTVED LELEDVLKAG YGGVKCVES GGPEPGVGCA GRGVITAINF LEEEGAYEDD LDFVFYDVLG DVVCGGFAMP IRENKAQEIY IVCSGEMMAM Y AANNISKG ...String: MAMRQCAIYG KGGIGKSTTT QNLVAALAEM GKKVMIVGCD PKADSTRLIL HSKAQNTIME MAAEAGTVED LELEDVLKAG YGGVKCVES GGPEPGVGCA GRGVITAINF LEEEGAYEDD LDFVFYDVLG DVVCGGFAMP IRENKAQEIY IVCSGEMMAM Y AANNISKG IVKYANSGSV RLGGLICNSR NTDREDELII ALANKLGTQM IHFVPRDNVV QRAEIRRMTV IEYDPKAKQA DE YRALARK VVDNKLLVIP NPITMDELEE LLMEFGIMEV EDESIVGKTA EEV |
-Experimental details
-Structure determination
Method | ![]() |
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Aggregation state | particle |
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Sample preparation
Concentration | 3.6 mg/mL | |||||||||
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Buffer | pH: 8 Component:
Details: Solutions were prepared and filtered immediately prior to the experiment. | |||||||||
Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Pretreatment - Type: PLASMA CLEANING | |||||||||
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 277 K / Instrument: HOMEMADE PLUNGER Details: Custom manual plunger. Greater than 95% humidity.. | |||||||||
Details | 3.6 mg/mL FeP Sample also contained 1.44 mg/mL MoFeP |
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Electron microscopy
Microscope | TFS KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD![]() |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Temperature | Min: 93.0 K / Max: 123.0 K |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 14903 / Average electron dose: 65.0 e/Å2 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - Chain ID: EF / Chain - Source name: PDB / Chain - Initial model type: experimental model |
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Refinement | Space: REAL / Protocol: RIGID BODY FIT |