Introduction

Helical reconstruction from electron cryomicrographs has become a routine technique for macromolecular structure determination of helical assemblies since the first days of Fourier-based three-dimensional image reconstruction. In the past decade, the single-particle technique has had an important impact on the advancement of helical reconstruction. Here, we present the software package SPRING that combines Fourier based symmetry analysis and real-space helical processing into a single workflow. One of the most time-consuming steps in helical reconstruction is the determination of the initial symmetry parameters. First, we propose a class-based helical reconstruction approach that enables the simultaneous exploration and evaluation of many symmetry combinations at low resolution. Second, multiple symmetry solutions can be further assessed and refined by single-particle based helical reconstruction using the correlation of simulated and experimental power spectra. Finally, the 3D structure can be determined to high resolution. In order to validate the procedure, we use the reference specimen Tobacco Mosaic Virus (TMV). After refinement of the helical symmetry, a total of 50,000 asymmetric units from two micrographs are sufficient to reconstruct a subnanometer 3D structure of TMV at 6.4Å resolution. Furthermore, we introduce the individual programs of the software and discuss enhancements of the helical reconstruction workflow. Thanks to its user-friendly interface and documentation, SPRING can be utilized by the novice as well as the expert user. In addition to the study of well-ordered helical structures, the development of a streamlined workflow for single-particle based helical reconstruction opens new possibilities to analyze specimens that are heterogeneous in symmetries.

Publications

  1. SPRING - an image processing package for single-particle based helical reconstruction from electron cryomicrographs.
    Cite this
    Desfosses A, Ciuffa R, Gutsche I, Sachse C, 2014-01-01 - Journal of structural biology

Credits

  1. Ambroise Desfosses
    Developer

    EMBL - European Molecular Biology Laboratory, Structural and Computational Biology Unit, Germany

  2. Rodolfo Ciuffa
    Developer

    EMBL - European Molecular Biology Laboratory, Structural and Computational Biology Unit, Germany

  3. Irina Gutsche
    Developer

    Univ. Grenoble Alpes, UVHCI, France

  4. Carsten Sachse
    Investigator

    EMBL - European Molecular Biology Laboratory, Structural and Computational Biology Unit, Germany

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Summary
AccessionBT000242
Tool TypeApplication
Category
PlatformsLinux/Unix
Technologies
User InterfaceTerminal Command Line
Download Count0
Country/RegionGermany
Submitted ByCarsten Sachse