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Osmolyte-mediated encapsulation of proteins inside MS2 viral capsids
Jeff E Glasgow1, Stacy L Capehart, Matthew B Francis
1Department of Chemistry, University of California, Berkeley, California 94720, USA.
Researchers developed two methods to encapsulate enzymes in bacteriophage MS2 viral capsids. Encapsulation minimally affected enzyme kinetics, offering a scalable approach for studying enzyme behavior in controlled nanometer-sized compartments.
Area of Science:
- Biotechnology
- Biophysics
- Molecular Engineering
Background:
- Enzyme encapsulation in nanocontainers can modulate activity in vitro and in vivo.
- Quantitative data on encapsulation effects in defined compartments is limited.
- Bacteriophage MS2 viral capsids offer a potential nanometer-sized compartment for enzyme encapsulation.
Purpose of the Study:
- To characterize two improved methods for encapsulating heterologous molecules within MS2 viral capsids.
- To assess the impact of encapsulation on enzyme kinetics.
- To provide a practical and scalable method for enzyme encapsulation studies.
Main Methods:
- Encapsulation via DNA oligomer tagging and MS2 coat protein reassembly, enhanced by trimethylamine-N-oxide.
- Encapsulation using expressed proteins with genetically encoded negatively charged peptide tags to induce capsid reassembly.
- Characterization of encapsulated alkaline phosphatase kinetics.
Main Results:
- Trimethylamine-N-oxide significantly increased yields in the DNA tagging method.
- Genetically encoded peptide tags resulted in high yields of protein-containing capsids.
- Encapsulated alkaline phosphatase exhibited similar K(m) and slightly lower k(cat) compared to the free enzyme.
Conclusions:
- Two viable methods for enzyme encapsulation in MS2 capsids were established.
- Encapsulation using genetically encoded tags minimally impacts enzyme kinetics.
- This approach offers a scalable platform for studying enzyme behavior in protein-based nanocontainers.
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