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

Journal of Structural Biology
M B ShermanM F Schmid

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.

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Citations

Jun 19, 1998·Current Opinion in Structural Biology·A McGough
Jul 5, 2002·Journal of Structural Biology·Yuyao LiangZ Hong Zhou
Jul 27, 2001·Journal of Structural Biology·F MetozM F Schmid
Nov 11, 1999·Journal of Molecular Biology·M B ShermanM F Schmid

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