High-resolution cryo-EM structure of the trypanosoma brucei ribosome: A case study

Amedee Des Georges, Yaser Hashem, Sarah N. Buss, Fabrice Jossinet, Qin Zhang, Hstau Y. Liao, Jie Fu, Amy Jobe, Robert A. Grassucci, Robert Langlois, Chandrajit Bajaj, Eric Westhof, Susan Madison-Antenucci, Joachim Frank

Research output: Chapter in Book/Report/Conference proceedingChapter


Single-particle cryo-electron microscopy has the immense advantage over crystallography in being able to image frozen-hydrated biological complexes in their “native” state, in solution. For years the ribosome has been the benchmark sample for particles without symmetry. It has witnessed steady improvement in resolution from the very first single-particle 3D reconstruction to today’s reconstructions at near-atomic resolution. In this study, we describe the different steps of sample preparation, data collection, data processing, and modeling that led to the 5Å structure of the T. brucei ribosome [32]. A local resolution estimation demonstrates the extent to which resolution can be anisotropic and pinpoints regions of higher heterogeneity or structural flexibility. This study also shows an example of misuse of spatial frequency filters leading to overfitting of the data and the artifacts that can be observed in the resulting density map.

Original languageEnglish (US)
Title of host publicationApplied and Numerical Harmonic Analysis
PublisherSpringer International Publishing
Number of pages36
StatePublished - 2014

Publication series

NameApplied and Numerical Harmonic Analysis
ISSN (Print)2296-5009
ISSN (Electronic)2296-5017


  • Contrast transfer function
  • Local resolution
  • Procyclic form
  • Reference projection
  • Wiener filter

ASJC Scopus subject areas

  • Applied Mathematics


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