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The phase separation underlying the pyrenoid-based microalgal Rubisco supercharger
Tobias Wunder1, Steven Le Hung Cheng1, Soak-Kuan Lai1
1School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore.
Nature Communications
|December 1, 2018
Summary
Researchers found that two proteins, Rubisco and EPYC1, can self-assemble into liquid droplets, mimicking the pyrenoid compartment essential for photosynthesis in microalgae. This discovery aids efforts to engineer more efficient crops.
Area of Science:
- Biochemistry
- Plant Biology
- Synthetic Biology
Background:
- Rubisco's slow and promiscuous nature limits photosynthetic efficiency.
- Carbon-concentrating mechanisms (CCMs) evolved to overcome Rubisco's limitations.
- In microalgae, Rubisco is sequestered in the pyrenoid, a chloroplast compartment.
Purpose of the Study:
- To investigate the in vitro self-assembly of Rubisco and EPYC1 into liquid droplets.
- To determine if these droplets mimic the functional pyrenoid compartment.
- To explore the potential for engineering CCMs in crop plants.
Main Methods:
- In vitro reconstitution assays using purified Rubisco and EPYC1 proteins.
- Phase separation and liquid droplet formation analysis.
- Functional assays of phase-separated Rubisco.
Main Results:
- Rubisco and EPYC1 are necessary and sufficient for phase separation into liquid droplets.
- Phase-separated Rubisco retains its enzymatic function.
- Droplet composition is dynamic with rapid component exchange.
- Variability in phase separation observed with heterologous and chimeric Rubiscos.
Conclusions:
- Rubisco and EPYC1 drive pyrenoid liquid-liquid phase separation in vitro.
- This in vitro system provides a platform to study pyrenoid biochemistry.
- Findings can guide synthetic biology efforts to engineer CCMs in crops.
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