Benchmarking tomographic acquisition schemes for high-resolution structural biology

Beata Turoňová, Wim J. H. Hagen, Martin Obr, Shyamal Mosalaganti, J. Wouter Beugelink, Christian E. Zimmerli, Hans-Georg Kräusslich and Martin Beck ()
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Beata Turoňová: Structure and Computational Biology Unit
Wim J. H. Hagen: Structure and Computational Biology Unit
Martin Obr: Universitätsklinikum Heidelberg
Shyamal Mosalaganti: Structure and Computational Biology Unit
J. Wouter Beugelink: Structure and Computational Biology Unit
Christian E. Zimmerli: Structure and Computational Biology Unit
Hans-Georg Kräusslich: Universitätsklinikum Heidelberg
Martin Beck: Structure and Computational Biology Unit

Nature Communications, 2020, vol. 11, issue 1, 1-9

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-14535-2

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