2.7 Å cryo-EM structure of vitrified M. musculus H-chain apoferritin from a compact 200 keV cryo-microscope.
Amino Acid Sequence
Animals
Apoferritins
/ chemistry
Cryoelectron Microscopy
/ instrumentation
Crystallography
Image Processing, Computer-Assisted
Imaging, Three-Dimensional
Mice
Models, Molecular
Protein Structure, Secondary
Protein Subunits
Single Molecule Imaging
/ instrumentation
Static Electricity
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
27
11
2019
accepted:
16
04
2020
entrez:
7
5
2020
pubmed:
7
5
2020
medline:
1
8
2020
Statut:
epublish
Résumé
Here we present the structure of mouse H-chain apoferritin at 2.7 Å (FSC = 0.143) solved by single particle cryogenic electron microscopy (cryo-EM) using a 200 kV device, the Thermo Fisher Glacios®. This is a compact, two-lens illumination system with a constant power objective lens, without any energy filters or aberration correctors, often thought of as a "screening cryo-microscope". Coulomb potential maps reveal clear densities for main chain carbonyl oxygens, residue side chains (including alternative conformations) and bound solvent molecules. We used a quasi-crystallographic reciprocal space approach to fit model coordinates to the experimental cryo-EM map. We argue that the advantages offered by (a) the high electronic and mechanical stability of the microscope, (b) the high emission stability and low beam energy spread of the high brightness Field Emission Gun (X-FEG), (c) direct electron detection technology and (d) particle-based Contrast Transfer Function (CTF) refinement have contributed to achieving high resolution. Overall, we show that basic electron optical settings for automated cryo-electron microscopy imaging can be used to determine structures approaching atomic resolution.
Identifiants
pubmed: 32374767
doi: 10.1371/journal.pone.0232540
pii: PONE-D-19-33016
pmc: PMC7202636
doi:
Substances chimiques
Protein Subunits
0
Apoferritins
9013-31-4
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0232540Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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