+Open data
-Basic information
Entry | Database: PDB / ID: 7yqm | ||||||
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Title | 2.9-angstrom cryo-EM structure of Ecoli malate synthase G | ||||||
Components | Malate synthase G | ||||||
Keywords | BIOSYNTHETIC PROTEIN / glyoxylate / malate / MSG / citric acid cycel / malate synthase G | ||||||
Function / homology | Function and homology information malate synthase / malate synthase activity / glyoxylate cycle / glyoxylate catabolic process / tricarboxylic acid cycle / magnesium ion binding / cytosol Similarity search - Function | ||||||
Biological species | Escherichia coli DH5[alpha] (bacteria) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.89 Å | ||||||
Authors | Wu, K.-P. / Wu, Y.-M. / Lu, Y.-C. | ||||||
Funding support | Taiwan, 1items
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Citation | Journal: J Struct Biol / Year: 2023 Title: Cryo-EM reveals the structure and dynamics of a 723-residue malate synthase G. Authors: Meng-Ru Ho / Yi-Ming Wu / Yen-Chen Lu / Tzu-Ping Ko / Kuen-Phon Wu / Abstract: Determination of sub-100 kDa (kDa) structures by cryo-electron microscopy (EM) is a longstanding but not straightforward goal. Here, we present a 2.9-Å cryo-EM structure of a 723-amino acid apo- ...Determination of sub-100 kDa (kDa) structures by cryo-electron microscopy (EM) is a longstanding but not straightforward goal. Here, we present a 2.9-Å cryo-EM structure of a 723-amino acid apo-form malate synthase G (MSG) from Escherichia coli. The cryo-EM structure of the 82-kDa MSG exhibits the same global folding as structures resolved by crystallography and nuclear magnetic resonance (NMR) spectroscopy, and the crystal and cryo-EM structures are indistinguishable. Analyses of MSG dynamics reveal consistent conformational flexibilities among the three experimental approaches, most notably that the α/β domain exhibits structural heterogeneity. We observed that sidechains of F453, L454, M629, and E630 residues involved in hosting the cofactor acetyl-CoA and substrate rotate differently between the cryo-EM apo-form and complex crystal structures. Our work demonstrates that the cryo-EM technique can be used to determine structures and conformational heterogeneity of sub-100 kDa biomolecules to a quality as high as that obtained from X-ray crystallography and NMR spectroscopy. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7yqm.cif.gz | 131.7 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7yqm.ent.gz | 100.7 KB | Display | PDB format |
PDBx/mmJSON format | 7yqm.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7yqm_validation.pdf.gz | 1.2 MB | Display | wwPDB validaton report |
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Full document | 7yqm_full_validation.pdf.gz | 1.2 MB | Display | |
Data in XML | 7yqm_validation.xml.gz | 36.8 KB | Display | |
Data in CIF | 7yqm_validation.cif.gz | 52.2 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/yq/7yqm ftp://data.pdbj.org/pub/pdb/validation_reports/yq/7yqm | HTTPS FTP |
-Related structure data
Related structure data | 34029MC 7yqnC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 80581.344 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli DH5[alpha] (bacteria) / Strain: K12 / Gene: glcB, glc, b2976, JW2943 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P37330, malate synthase |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: 2.9-angstrom cryo-EM structure of 723-aa malate synthase G Type: CELL / Entity ID: all / Source: RECOMBINANT |
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Source (natural) | Organism: Escherichia coli DH5[alpha] (bacteria) |
Source (recombinant) | Organism: Escherichia coli BL21(DE3) (bacteria) |
Buffer solution | pH: 7.6 |
Specimen | Conc.: 0.3 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 1800 nm / Nominal defocus min: 1000 nm |
Image recording | Electron dose: 51.2 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 QUANTUM (4k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.19.2_4158: / Classification: refinement | ||||||||||||||||||||||||
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EM software |
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CTF correction | Type: NONE | ||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 2.89 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 211582 / Algorithm: FOURIER SPACE / Symmetry type: POINT | ||||||||||||||||||||||||
Atomic model building | PDB-ID: 1P7T Pdb chain-ID: A / Accession code: 1P7T / Source name: PDB / Type: experimental model | ||||||||||||||||||||||||
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