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Structure and function of the AAA+ protein CbbX, a red-type Rubisco activase

Nature volume 479, pages 194–199 (2011)Cite this article

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Abstract

Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyses the fixation of atmospheric CO2 in photosynthesis, but tends to form inactive complexes with its substrate ribulose 1,5-bisphosphate (RuBP). In plants, Rubisco is reactivated by the AAA+ (ATPases associated with various cellular activities) protein Rubisco activase (Rca), but no such protein is known for the Rubisco of red algae. Here we identify the protein CbbX as an activase of red-type Rubisco. The 3.0-Å crystal structure of unassembled CbbX from Rhodobacter sphaeroides revealed an AAA+ protein architecture. Electron microscopy and biochemical analysis showed that ATP and RuBP must bind to convert CbbX into functionally active, hexameric rings. The CbbX ATPase is strongly stimulated by RuBP and Rubisco. Mutational analysis suggests that CbbX functions by transiently pulling the carboxy-terminal peptide of the Rubisco large subunit into the hexamer pore, resulting in the release of the inhibitory RuBP. Understanding Rubisco activation may facilitate efforts to improve CO2 uptake and biomass production by photosynthetic organisms.

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Figure 1: CbbX functions as a Rubisco activase.
Figure 2: Negative-stain electron microscopy of CbbX.
Figure 3: Crystal structure and hexamer model of CbbX.
Figure 4: Structural and functional analysis of CbbX mechanism.
Figure 5: Model of Rubisco activation by CbbX.

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Protein Data Bank

Data deposits

Coordinates and structure factor amplitudes for CbbX crystal structures are deposited in the Protein Data Bank (PDB) under accession codes 3SYL and 3SYK; the hexamer model and the electron microscopy density are deposited in the PDB under accession code 3ZUH and in the Electron Microscopy Database (http://www.ebi.ac.uk/pdbe/emdb/) under accession code EMD-1932, respectively.

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Acknowledgements

We thank S. Kaplan for providing the R. sphaeroides strain 2.4.1, S. Whitney for providing the pHue protein expression system, and R. Lange and N. Wischnewski for technical assistance. Support by the Max Planck Institute of Biochemistry (MPIB) Core Facility, the MPIB Crystallization Facility and the Joint Structural Biology Group staff at the European Synchrotron Radiation Facility beamlines is gratefully acknowledged. We thank the Deutsche Forschungsgemeinschaft (DFG) (SFB 594; DFG grant WE4628/1 to P.W.) and the Körber Foundation for financial support.

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  1. Department of Cellular Biochemistry, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany,

    Oliver Mueller-Cajar, Mathias Stotz, F. Ulrich Hartl, Andreas Bracher & Manajit Hayer-Hartl

  2. Department of Biochemistry, Gene Center Munich, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strasse 25, 81377 Munich, Germany,

    Petra Wendler

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Contributions

O.M.-C. designed and performed all the biochemical experiments. O.M.-C., M.S. and A.B. obtained the CbbX crystals and solved the structure. P.W. performed the electron microscopy and three-dimensional image analysis. All authors contributed to data interpretation and manuscript preparation. O.M.-C., A.B., F.U.H. and M.H.-H. wrote the manuscript.

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Correspondence to Andreas Bracher or Manajit Hayer-Hartl.

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Mueller-Cajar, O., Stotz, M., Wendler, P. et al. Structure and function of the AAA+ protein CbbX, a red-type Rubisco activase. Nature 479, 194–199 (2011). https://doi.org/10.1038/nature10568

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