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TitleVisualizing Individual RuBisCO and Its Assembly into Carboxysomes in Marine Cyanobacteria by Cryo-Electron Tomography.
Journal, issue, pagesJ Mol Biol, Vol. 430, Issue 21, Page 4156-4167, Year 2018
Publish dateOct 19, 2018
AuthorsWei Dai / Muyuan Chen / Christopher Myers / Steven J Ludtke / B Montgomery Pettitt / Jonathan A King / Michael F Schmid / Wah Chiu /
PubMed AbstractCyanobacteria are photosynthetic organisms responsible for ~25% of the organic carbon fixation on earth. A key step in carbon fixation is catalyzed by ribulose bisphosphate carboxylase/oxygenase ...Cyanobacteria are photosynthetic organisms responsible for ~25% of the organic carbon fixation on earth. A key step in carbon fixation is catalyzed by ribulose bisphosphate carboxylase/oxygenase (RuBisCO), the most abundant enzyme in the biosphere. Applying Zernike phase-contrast electron cryo-tomography and automated annotation, we identified individual RuBisCO molecules and their assembly intermediates leading to the formation of carboxysomes inside Syn5 cyanophage infected cyanobacteria Synechococcus sp. WH8109 cells. Surprisingly, more RuBisCO molecules were found to be present as cytosolic free-standing complexes or clusters than as packaged assemblies inside carboxysomes. Cytosolic RuBisCO clusters and partially assembled carboxysomes identified in the cell tomograms support a concurrent assembly model involving both the protein shell and the enclosed RuBisCO. In mature carboxysomes, RuBisCO is neither randomly nor strictly icosahedrally packed within protein shells of variable sizes. A time-averaged molecular dynamics simulation showed a semi-liquid probability distribution of the RuBisCO in carboxysomes and correlated well with carboxysome subtomogram averages. Our structural observations reveal the various stages of RuBisCO assemblies, which could be important for understanding cellular function.
External linksJ Mol Biol / PubMed:30138616 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution40.0 Å
Structure data

EMDB-7854:
Carboxysome subtomogram averaging of size group 104 nm
Method: EM (subtomogram averaging) / Resolution: 40.0 Å

EMDB-7855:
Carboxysome subtomogram averaging of size group 106 nm
Method: EM (subtomogram averaging) / Resolution: 40.0 Å

EMDB-7856:
Carboxysome subtomogram averaging of size group 108 nm
Method: EM (subtomogram averaging) / Resolution: 40.0 Å

EMDB-7857:
Carboxysome subtomogram averaging of size group 110 nm
Method: EM (subtomogram averaging) / Resolution: 40.0 Å

Source
  • Synechococcus sp. WH 8109 (bacteria)

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