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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.

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Summary
This summary is machine-generated.

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.

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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.