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Rubisco forms a lattice inside alpha-carboxysomes

Metskas, Lauren Ann and Ortega, Davi and Oltrogge, Luke M. and Blikstad, Cecilia and Lovejoy, Derik R. and Laughlin, Thomas G. and Savage, David F. and Jensen, Grant J. (2022) Rubisco forms a lattice inside alpha-carboxysomes. Nature Communications, 13 (1). Art. No. 4863. ISSN 2041-1723. doi:10.1038/s41467-022-32584-7.

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Despite the importance of microcompartments in prokaryotic biology and bioengineering, structural heterogeneity has prevented a complete understanding of their architecture, ultrastructure, and spatial organization. Here, we employ cryo-electron tomography to image α-carboxysomes, a pseudo-icosahedral microcompartment responsible for carbon fixation. We have solved a high-resolution subtomogram average of the Rubisco cargo inside the carboxysome, and determined the arrangement of the enzyme. We find that the H. neapolitanus Rubisco polymerizes in vivo, mediated by the small Rubisco subunit. These fibrils can further pack to form a lattice with six-fold pseudo-symmetry. This arrangement preserves freedom of motion and accessibility around the Rubisco active site and the binding sites for two other carboxysome proteins, CsoSCA (a carbonic anhydrase) and the disordered CsoS2, even at Rubisco concentrations exceeding 800 μM. This characterization of Rubisco cargo inside the α-carboxysome provides insight into the balance between order and disorder in microcompartment organization.

Item Type:Article
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URLURL TypeDescription ReadCube access ItemDiscussion paper
Metskas, Lauren Ann0000-0002-8073-6960
Ortega, Davi0000-0002-8344-2335
Oltrogge, Luke M.0000-0001-5716-9980
Blikstad, Cecilia0000-0001-5740-926X
Laughlin, Thomas G.0000-0001-8943-1330
Savage, David F.0000-0003-0042-2257
Jensen, Grant J.0000-0003-1556-4864
Additional Information:Cryo-electron microscopy was done in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech, and subtomogram alignment and averaging used the Caltech Resnick High Performance Computing Center. We thank S. Chen and A. Malyutin for assistance with tomography data collection, A. Burt for pseudocode to transition between Dynamo and Relion software packages, and A. Pranger for assistance reconstructing cellular tomograms. This work was supported by a Ruth L. Kirschstein NRSA Individual Postdoctoral Fellowship F32 1F32GM135994-01 to L.A.M., NIH R01GM129241 to D.F.S., and NIH R01 AI127401 to G.J.J.
Funding AgencyGrant Number
NIHF32 1F32GM135994-01
NIHR01 AI127401
Issue or Number:1
Record Number:CaltechAUTHORS:20220829-467858200
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:116620
Deposited By: Melissa Ray
Deposited On:02 Sep 2022 17:22
Last Modified:02 Sep 2022 17:22

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