2.7 A cryo-EM structure of vitrified M. musculus H-chain apoferritin from 200 keV \"screening microscope\"

Hamdi, F.; Tueting, C.; Semchonok, D. A.; Kyrilis, F. L.; Meister, A.; Skalidis, I.; Schmidt, L.; Parthier, C.; Stubbs, M. T.; Kastritis, P. L.

doi:10.1101/738724

bioRxiv · 10.1101/738724

2.7 A cryo-EM structure of vitrified M. musculus H-chain apoferritin from 200 keV \"screening microscope\"

Hamdi, F.·Tueting, C.·Semchonok, D. A.·Kyrilis, F. L.·Meister, A.·Skalidis, I.·Schmidt, L.·Parthier, C.·Stubbs, M. T.·Kastritis, P. L.

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Abstract

Here we present the structure of mouse H-chain apoferritin at 2.7 [A] (FSC=0.143) solved by single particle cryogenic electron microscopy (cryo-EM) using a 200 kV device. Data were collected using a compact, two-lens illumination system with a constant power objective lens, without the use of energy filters or aberration correctors. Coulomb potential maps reveal clear densities for main chain carbonyl oxygens, residue side chains (including alternative conformations) and bound solvent molecules. 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 resolution close to the Rayleigh limit. Overall, we show that basic electron optical settings for automated cryo-electron microscopy imaging, widely thought of as a "screening cryo-microscope", can be used to determine structures approaching atomic resolution. HighlightsO_LIThe 2.7 [A] structure of mouse apoferritin was solved using a 200 keV screening cryo-microscope C_LIO_LIThe apoferritin reconstruction was resolved without an energy filter, aberration correctors, or constant-power condenser lenses C_LIO_LIComparison to available crystallographic and cryo-EM structures from high-end cryo-microscopes demonstrates consistency in resolved water molecules, metals and side chain orientations C_LIO_LIAlthough radiation damage is more prominent at 200 keV compared to 300 keV, this type of instrumentation is more accessible to research laboratories due to its compactness and simplicity C_LI

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Hamdi, F., Tueting, C., Semchonok, D. A., Kyrilis, F. L., Meister, A., Skalidis, I., Schmidt, L., Parthier, C., Stubbs, M. T., Kastritis, P. L.. 2019-08-20. 2.7 A cryo-EM structure of vitrified M. musculus H-chain apoferritin from 200 keV \"screening microscope\". https://doi.org/10.1101/738724

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