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TitleSub-2 Angstrom resolution structure determination using single-particle cryo-EM at 200 keV.
Journal, issue, pagesJ Struct Biol X, Vol. 4, Page 100020, Year 2020
Publish dateFeb 28, 2020
AuthorsMengyu Wu / Gabriel C Lander / Mark A Herzik /
PubMed AbstractAlthough the advent of direct electron detectors (DEDs) and software developments have enabled the routine use of single-particle cryogenic electron microscopy (cryo-EM) for structure determination ...Although the advent of direct electron detectors (DEDs) and software developments have enabled the routine use of single-particle cryogenic electron microscopy (cryo-EM) for structure determination of well-behaved specimens to high-resolution, there nonetheless remains a discrepancy between the resolutions attained for biological specimens and the information limits of modern transmission electron microscopes (TEMs). Instruments operating at 300 kV equipped with DEDs are the current paradigm for high-resolution single-particle cryo-EM, while 200 kV TEMs remain comparatively underutilized for purposes beyond sample screening. Here, we expand upon our prior work and demonstrate that one such 200 kV microscope, the Talos Arctica, equipped with a K2 DED is capable of determining structures of macromolecules to as high as ∼1.7 Å resolution. At this resolution, ordered water molecules are readily assigned and holes in aromatic residues can be clearly distinguished in the reconstructions. This work emphasizes the utility of 200 kV electrons for high-resolution single-particle cryo-EM and applications such as structure-based drug design.
External linksJ Struct Biol X / PubMed:32647824 / PubMed Central
MethodsEM (single particle)
Resolution1.75 - 2.13 Å
Structure data

21023

6v20

EMDB-21023: Single-particle cryo-EM reconstruction of rabbit muscle aldolase using 200 keV
PDB-6v20: Rabbit muscle aldolase determined using single-particle cryo-EM at 200 keV
Method: EM (single particle) / Resolution: 2.13 Å

21024

6v21

EMDB-21024: Single-particle cryo-EM reconstruction of mouse heavy chain apoferritin at 200 keV
PDB-6v21: Mouse heavy chain apoferritin determined using single-particle cryo-EM at 200 keV
Method: EM (single particle) / Resolution: 1.75 Å

Chemicals

ChemComp-HOH:
WATER / Water

Source
  • oryctolagus cuniculus (rabbit)
  • mus musculus (house mouse)
KeywordsLYASE / homo-4-mer / D-fructose / glycolysis / METAL BINDING PROTEIN / OXIDOREDUCTASE / homo-24-mer / storage / globular

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