Benchmarking tomographic acquisition schemes for high-resolution structural biology

B. Turoňová, W.J.H. Hagen, M. Obr, S. Mosalaganti, J.W. Beugelink, C.E. Zimmerli, H.G. Kräusslich, M. Beck, Nature Communications 11 (2020).

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Journal Article | Published | English
Author
Turoňová, Beata; Hagen, Wim J.H.; Obr, MartinIST Austria; Mosalaganti, Shyamal; Beugelink, J. Wouter; Zimmerli, Christian E.; Kräusslich, Hans Georg; Beck, Martin
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Abstract
Cryo electron tomography with subsequent subtomogram averaging is a powerful technique to structurally analyze macromolecular complexes in their native context. Although close to atomic resolution in principle can be obtained, it is not clear how individual experimental parameters contribute to the attainable resolution. Here, we have used immature HIV-1 lattice as a benchmarking sample to optimize the attainable resolution for subtomogram averaging. We systematically tested various experimental parameters such as the order of projections, different angular increments and the use of the Volta phase plate. We find that although any of the prominently used acquisition schemes is sufficient to obtain subnanometer resolution, dose-symmetric acquisition provides considerably better outcome. We discuss our findings in order to provide guidance for data acquisition. Our data is publicly available and might be used to further develop processing routines.
Publishing Year
Date Published
2020-02-13
Journal Title
Nature communications
Volume
11
Article Number
876
eISSN
IST-REx-ID

Cite this

Turoňová B, Hagen WJH, Obr M, et al. Benchmarking tomographic acquisition schemes for high-resolution structural biology. Nature communications. 2020;11. doi:10.1038/s41467-020-14535-2
Turoňová, B., Hagen, W. J. H., Obr, M., Mosalaganti, S., Beugelink, J. W., Zimmerli, C. E., … Beck, M. (2020). Benchmarking tomographic acquisition schemes for high-resolution structural biology. Nature Communications, 11. https://doi.org/10.1038/s41467-020-14535-2
Turoňová, Beata, Wim J.H. Hagen, Martin Obr, Shyamal Mosalaganti, J. Wouter Beugelink, Christian E. Zimmerli, Hans Georg Kräusslich, and Martin Beck. “Benchmarking Tomographic Acquisition Schemes for High-Resolution Structural Biology.” Nature Communications 11 (2020). https://doi.org/10.1038/s41467-020-14535-2.
B. Turoňová et al., “Benchmarking tomographic acquisition schemes for high-resolution structural biology,” Nature communications, vol. 11, 2020.
Turoňová B, Hagen WJH, Obr M, Mosalaganti S, Beugelink JW, Zimmerli CE, Kräusslich HG, Beck M. 2020. Benchmarking tomographic acquisition schemes for high-resolution structural biology. Nature communications. 11.
Turoňová, Beata, et al. “Benchmarking Tomographic Acquisition Schemes for High-Resolution Structural Biology.” Nature Communications, vol. 11, 876, Springer Nature, 2020, doi:10.1038/s41467-020-14535-2.
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