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TitleCost-benefit analysis of cryogenic electron tomography subtomogram averaging of chaperonin MmCpn at near atomic resolution.
Journal, issue, pagesStructure, Vol. 33, Issue 2, Page 372-380.e2, Year 2025
Publish dateFeb 6, 2025
AuthorsYanyan Zhao / Michael F Schmid / Wah Chiu /
PubMed AbstractCryogenic electron microscopy single particle analysis (cryoEM-SPA) has evolved into a routine approach for determining macromolecule structures to near-atomic resolution. Cryogenic electron ...Cryogenic electron microscopy single particle analysis (cryoEM-SPA) has evolved into a routine approach for determining macromolecule structures to near-atomic resolution. Cryogenic electron tomography subtomogram averaging (cryoET-STA) toward a similar resolution, in contrast, is still under active development. Here, we use the archeal chaperonin MmCpn as a model macromolecule to quantitatively investigate the resolution limiting factors of cryoET-STA in terms of cumulative electron dose, ice thickness, subtomogram numbers, and tilt angle ranges. By delineating the feasibility and experimental factors of attaining near atomic resolution structure with cryoET-STA, especially the effect of electron damage through the tilt series and inelastic scattering at various ice thickness, we encourage a customized tilt series collection strategy for efficient throughput. This study provides a biophysical basis for the application of cryoET-STA (for highly symmetric molecules like MmCpn) toward high resolution and the rationales in using cryoET-STA to achieve an efficient outcome at the desired resolution.
External linksStructure / PubMed:39644888 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution2.56 - 3.6 Å
Structure data

45793

9cpk

EMDB-45793: Closed state MmCpn under ATP/AlFx condition by subtomogram averaging
PDB-9cpk: D8 symmetry reconstruction of MmCPn in closed state
Method: EM (subtomogram averaging) / Resolution: 2.56 Å

EMDB-45794: MmCpn in closed state wit D9 symmetry
Method: EM (subtomogram averaging) / Resolution: 3.6 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-AF3:
ALUMINUM FLUORIDE

ChemComp-K:
Unknown entry

ChemComp-HOH:
WATER

Source
  • Archaea (unknown)
  • methanococcus maripaludis (archaea)
KeywordsCHAPERONE / Archaea chaperonin in closed state with ATP/AlFx

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