Identification of a carbonic anhydrase–Rubisco complex within the alpha-carboxysome
Abstract
Carboxysomes are proteinaceous organelles that encapsulate key enzymes of CO 2 fixation—Rubisco and carbonic anhydrase—and are the centerpiece of the bacterial CO 2 concentrating mechanism (CCM). In the CCM, actively accumulated cytosolic bicarbonate diffuses into the carboxysome and is converted to CO 2 by carbonic anhydrase, producing a high CO 2 concentration near Rubisco and ensuring efficient carboxylation. Self-assembly of the α-carboxysome is orchestrated by the intrinsically disordered scaffolding protein, CsoS2, which interacts with both Rubisco and carboxysomal shell proteins, but it is unknown how the carbonic anhydrase, CsoSCA, is incorporated into the α-carboxysome. Here, we present the structural basis of carbonic anhydrase encapsulation into α-carboxysomes from Halothiobacillus neapolitanus . We find that CsoSCA interacts directly with Rubisco via an intrinsically disordered N-terminal domain. A 1.98 Å single-particle cryoelectron microscopy structure of Rubisco in complex with this peptide reveals that CsoSCA binding is predominantly mediated by a network of hydrogen bonds. CsoSCA's binding site overlaps with that of CsoS2, but the two proteins utilize substantially different motifs and modes of binding, revealing a plasticity of the Rubisco binding site. Our results advance the understanding of carboxysome biogenesis and highlight the importance of Rubisco, not only as an enzyme but alsomore »
- Authors:
-
[1];
[2];
[2];
[2];
[3];
[2];
[4];
[5];
[6]
- Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, Department of Chemistry - Ångström Laboratory, Uppsala University, Uppsala 75120, Sweden
- Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720
- Department of Chemistry, University of California, Berkeley, CA 94720
- Department of Chemistry - Ångström Laboratory, Uppsala University, Uppsala 75120, Sweden
- California Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720
- Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, HHMI, University of California, Berkeley, CA 94720
- Publication Date:
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 2203163
- Grant/Contract Number:
- SC00016240
- Resource Type:
- Published Article
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America
- Additional Journal Information:
- Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 120 Journal Issue: 43; Journal ID: ISSN 0027-8424
- Publisher:
- Proceedings of the National Academy of Sciences
- Country of Publication:
- United States
- Language:
- English
Citation Formats
Blikstad, Cecilia, Dugan, Eli J., Laughlin, Thomas G., Turnšek, Julia B., Liu, Mira D., Shoemaker, Sophie R., Vogiatzi, Nikoleta, Remis, Jonathan P., and Savage, David F. Identification of a carbonic anhydrase–Rubisco complex within the alpha-carboxysome. United States: N. p., 2023.
Web. doi:10.1073/pnas.2308600120.
Blikstad, Cecilia, Dugan, Eli J., Laughlin, Thomas G., Turnšek, Julia B., Liu, Mira D., Shoemaker, Sophie R., Vogiatzi, Nikoleta, Remis, Jonathan P., & Savage, David F. Identification of a carbonic anhydrase–Rubisco complex within the alpha-carboxysome. United States. https://doi.org/10.1073/pnas.2308600120
Blikstad, Cecilia, Dugan, Eli J., Laughlin, Thomas G., Turnšek, Julia B., Liu, Mira D., Shoemaker, Sophie R., Vogiatzi, Nikoleta, Remis, Jonathan P., and Savage, David F. Fri .
"Identification of a carbonic anhydrase–Rubisco complex within the alpha-carboxysome". United States. https://doi.org/10.1073/pnas.2308600120.
@article{osti_2203163,
title = {Identification of a carbonic anhydrase–Rubisco complex within the alpha-carboxysome},
author = {Blikstad, Cecilia and Dugan, Eli J. and Laughlin, Thomas G. and Turnšek, Julia B. and Liu, Mira D. and Shoemaker, Sophie R. and Vogiatzi, Nikoleta and Remis, Jonathan P. and Savage, David F.},
abstractNote = {Carboxysomes are proteinaceous organelles that encapsulate key enzymes of CO 2 fixation—Rubisco and carbonic anhydrase—and are the centerpiece of the bacterial CO 2 concentrating mechanism (CCM). In the CCM, actively accumulated cytosolic bicarbonate diffuses into the carboxysome and is converted to CO 2 by carbonic anhydrase, producing a high CO 2 concentration near Rubisco and ensuring efficient carboxylation. Self-assembly of the α-carboxysome is orchestrated by the intrinsically disordered scaffolding protein, CsoS2, which interacts with both Rubisco and carboxysomal shell proteins, but it is unknown how the carbonic anhydrase, CsoSCA, is incorporated into the α-carboxysome. Here, we present the structural basis of carbonic anhydrase encapsulation into α-carboxysomes from Halothiobacillus neapolitanus . We find that CsoSCA interacts directly with Rubisco via an intrinsically disordered N-terminal domain. A 1.98 Å single-particle cryoelectron microscopy structure of Rubisco in complex with this peptide reveals that CsoSCA binding is predominantly mediated by a network of hydrogen bonds. CsoSCA's binding site overlaps with that of CsoS2, but the two proteins utilize substantially different motifs and modes of binding, revealing a plasticity of the Rubisco binding site. Our results advance the understanding of carboxysome biogenesis and highlight the importance of Rubisco, not only as an enzyme but also as a central hub for mediating assembly through protein interactions.},
doi = {10.1073/pnas.2308600120},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 43,
volume = 120,
place = {United States},
year = {Fri Oct 20 00:00:00 EDT 2023},
month = {Fri Oct 20 00:00:00 EDT 2023}
}
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