A strategy for electron tomographic data collection and crystallographic reconstruction of biological bundles

Michael B. Sherman, Joanita Jakana, Shujun Sun, Paul Matsudaira, Wah Chiu, Michael F. Schmid

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Structures of highly ordered biological bundles have unique features which call for special experimental and computational methods in electron cryomicroscopy. They can be considered as three-dimensional quasi-crystals and reconstructed using a crystallographic approach. However, they are neither 'infinitely' large with respect to the borders of the bundle, nor are they a single unit cell in thickness along the viewing direction. Also, because of their shape, bundles do not generally have a preferred azimuthal orientation, which poses challenges for orientation estimation and refinement. We developed a strategy for recording and processing electron cryomicroscopic images that differs from classical two-dimensional crystalline reconstruction techniques. These developments allowed us to merge data from tomographic tilt series of ice-embedded acrosomal bundles. The goal is to determine accurately amplitudes and phases at the diffraction maxima in terms of hkl indices, and compute a three-dimensional map from the diffraction data.

Original languageEnglish (US)
Pages (from-to)245-256
Number of pages12
JournalJournal of Structural Biology
Volume120
Issue number3
DOIs
StatePublished - Dec 1997
Externally publishedYes

Keywords

  • Acrosomal bundles
  • Actin
  • Electron cryomicroscopy
  • Electron crystallography
  • Profile fitting
  • Scruin
  • Tilt series imaging
  • Tomography

ASJC Scopus subject areas

  • Structural Biology

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