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TitleCryo-EM reveals the structure and dynamics of a 723-residue malate synthase G.
Journal, issue, pagesJ Struct Biol, Vol. 215, Issue 2, Page 107958, Year 2023
Publish dateMar 28, 2023
AuthorsMeng-Ru Ho / Yi-Ming Wu / Yen-Chen Lu / Tzu-Ping Ko / Kuen-Phon Wu /
PubMed AbstractDetermination 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.
External linksJ Struct Biol / PubMed:36997036
MethodsEM (single particle) / X-ray diffraction
Resolution1.6 - 4.14 Å
Structure data

34029

7yqm

EMDB-34029, PDB-7yqm:
2.9-angstrom cryo-EM structure of Ecoli malate synthase G
Method: EM (single particle) / Resolution: 2.89 Å

EMDB-34030: Cryo-EM map of a dimeric form of Ecoli Malate Synthase G (MSG)
Method: EM (single particle) / Resolution: 4.14 Å

PDB-7yqn:
Crystal structure of Ecoli malate synthase G
Method: X-RAY DIFFRACTION / Resolution: 1.6 Å

Chemicals

ChemComp-CL:
Unknown entry

ChemComp-NA:
Unknown entry

ChemComp-BME:
BETA-MERCAPTOETHANOL

ChemComp-GOL:
GLYCEROL

ChemComp-HOH:
WATER

Source
  • escherichia coli dh5[alpha] (bacteria)
KeywordsBIOSYNTHETIC PROTEIN / glyoxylate / malate / MSG / citric acid cycel / malate synthase G

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