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Recombinant Fusion Proteins with Embedded Sensing Functions as Versatile Tools for Protocell Development
Bornita Deb1, Adriana LaVopa2, Emma McDougal1,3
1Department of Chemical Engineering, University of Florida, Gainesville, Florida 32611, United States.
Biomacromolecules
|December 3, 2024
Summary
Researchers created synthetic protein vesicles that sense molecules like rapamycin. These sensory globular protein vesicles (GPVs) self-assemble and aggregate upon detecting specific signals, mimicking cellular functions.
Area of Science:
- Biotechnology
- Synthetic Biology
- Biochemistry
Background:
- Cellular sensory capabilities are essential for environmental interaction.
- Mimicking cellular sensing in synthetic systems is a key challenge.
- Protein-based self-assembly offers a route to construct functional synthetic compartments.
Purpose of the Study:
- To develop sensory globular protein vesicles (GPVs) capable of detecting specific signaling molecules.
- To engineer GPVs using self-assembling recombinant fusion proteins.
- To investigate the mechanism of sensory response and vesicle aggregation.
Main Methods:
- Genetic fusion of sensory domains (FKBP/FRB) with fluorescent proteins and leucine zippers.
- Self-assembly of amphiphilic building blocks into vesicles in aqueous conditions.
- Utilizing high-affinity leucine zipper binding and elastin-like polypeptide (ELP) properties for vesicle formation.
- Monitoring intervesicle aggregation via colocalization and statistical analysis upon rapamycin addition.
Main Results:
- Successful self-assembly of sensory globular protein vesicles (GPVs) from recombinant fusion proteins.
- Demonstrated sensory function through FKBP-FRB complex formation induced by rapamycin.
- Observed time- and concentration-dependent intervesicle aggregation triggered by rapamycin binding.
- Confirmed aggregation through colocalization studies and statistical validation.
Conclusions:
- Developed a novel system of sensory globular protein vesicles (GPVs) for molecular detection.
- The GPV system provides a platform for understanding protein vesicle sensing mechanisms.
- Offers a foundation for using GPVs as models for cellular processes in synthetic cells.
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