Sub-2 Angstrom resolution structure determination using single-particle cryo-EM at 200 keV

[Display omitted] •200 kV instruments enable structure determination of complexes to better than 2 Å resolution.•Regions of apoferritin structure resolved to ∼1.6 Å resolution by single-particle cryo-EM.•∼2.1 Å structure of ∼150 kDa rabbit muscle aldolase.•Resolution-limiting aberrations can now be...

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Veröffentlicht in:Journal of structural biology. X 2020-01, Vol.4, p.100020-100020, Article 100020
Hauptverfasser: Wu, Mengyu, Lander, Gabriel C., Herzik, Mark A.
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Sprache:eng
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Zusammenfassung:[Display omitted] •200 kV instruments enable structure determination of complexes to better than 2 Å resolution.•Regions of apoferritin structure resolved to ∼1.6 Å resolution by single-particle cryo-EM.•∼2.1 Å structure of ∼150 kDa rabbit muscle aldolase.•Resolution-limiting aberrations can now be estimated and corrected in silico. 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.
ISSN:2590-1524
2590-1524
DOI:10.1016/j.yjsbx.2020.100020