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TitleBenchmarking tomographic acquisition schemes for high-resolution structural biology.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 876, Year 2020
Publish dateFeb 13, 2020
AuthorsBeata Turoňová / Wim J H Hagen / Martin Obr / Shyamal Mosalaganti / J Wouter Beugelink / Christian E Zimmerli / Hans-Georg Kräusslich / Martin Beck /
PubMed AbstractCryo 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 ...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.
External linksNat Commun / PubMed:32054835 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution4.2 Å
Structure data

EMDB-10207:
A 4.2 Angstrom structure of HIV-1 CA-SP1
Method: EM (subtomogram averaging) / Resolution: 4.2 Å

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