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In Vitro Assembly of Diverse Bacterial Microcompartment Shell Architectures
Andrew R Hagen1, Jefferson S Plegaria2, Nancy Sloan1
1Environmental Genomics and Systems Biology and Molecular Biophysics and Integrated Bioimaging Divisions , Lawrence Berkeley National Laboratory , 1 Cyclotron Road , Berkeley , California 94720 , United States.
Nano Letters
|October 23, 2018
Summary
Researchers developed a new in vitro method to assemble bacterial microcompartment (BMC) architectures, including shells and nanotubes. This breakthrough enables precise engineering of BMCs for applications in nanoreactors and molecular scaffolds.
Area of Science:
- Biochemistry
- Synthetic Biology
- Nanotechnology
Background:
- Bacterial microcompartments (BMCs) are protein-bound organelles that enhance metabolic pathway efficiency.
- Their self-assembling nature makes them promising for bioengineering, but in vivo assembly testing is challenging.
- Existing methods limit the potential of BMCs in creating novel nanostructures.
Purpose of the Study:
- To develop a novel in vitro method for assembling defined bacterial microcompartment (BMC) architectures.
- To overcome limitations of in vivo expression for BMC engineering and functional testing.
- To demonstrate the utility of in vitro assembly for creating engineered BMC-based nanoreactors.
Main Methods:
- A SUMO (small ubiquitin-like modifier) tag was used as a "protecting group" to control BMC protein self-assembly in vivo.
- Shell proteins were concentrated in vitro after SUMO tag removal by a cognate protease.
- Mixing of constituent shell proteins in vitro facilitated self-assembly into various BMC architectures.
Main Results:
- Successfully achieved in vitro self-assembly of metabolosome shells, carboxysome shells, and BMC protein-based nanotubes.
- Engineered a metabolosome shell with a hyper-basic luminal surface for charge-complementary cargo encapsulation.
- Demonstrated the first use of charge complementarity for encapsulating diverse biotic and abiotic cargos within BMC shells.
Conclusions:
- A versatile in vitro method for assembling natural and engineered BMC architectures was established.
- This method significantly advances the bioengineering potential of BMCs for creating custom nanoreactors.
- The developed technique offers a broadly applicable platform for designing and constructing novel protein-based nanomaterials.
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