Structure of Halothiobacillus neapolitanus carboxysomes by cryo-electron tomography

J Mol Biol. 2006 Dec 1;364(3):526-35. doi: 10.1016/j.jmb.2006.09.024. Epub 2006 Sep 14.

Abstract

Carboxysomes are polyhedral bodies consisting of a proteinaceous shell filled with ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO). They are found in the cytoplasm of all cyanobacteria and some chemoautotrophic bacteria. Previous studies of Halothiobacillus neapolitanus and Nitrobacter agilis carboxysomes suggest that the structures are either icosahedral or dodecahedral. To determine the protein shell structure more definitively, purified H. neapolitanus carboxysomes were re-examined by cryo-electron tomography and scanning transmission electron microscopy (STEM). Due to the limited tilt angles in the electron microscope, the tomographic reconstructions are distorted. Corrections were made in the 3D orientation searching and averaging of the computationally extracted carboxysomes to minimize the missing data effects. It was found that H. neapolitanus carboxysomes vary widely in size and mass as shown by cryo-electron tomography and STEM mass measurements, respectively. We have aligned and averaged carboxysomes in several size classes from the 3D tomographic reconstruction by methods that are not model-biased. The averages reveal icosahedral symmetry of the shell, but not of the density inside it, for all the size classes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cryoelectron Microscopy
  • Halothiobacillus / metabolism*
  • Image Processing, Computer-Assisted
  • Imaging, Three-Dimensional
  • Microscopy, Electron, Scanning Transmission
  • Ribulose-Bisphosphate Carboxylase / chemistry*

Substances

  • Ribulose-Bisphosphate Carboxylase