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Secretory vesicles, also known as dense core vesicles (DCVs), are membrane-bound vesicles that transport secretory proteins, such as hormones or neurotransmitters. Regulated secretory vesicles transport proteins from the trans-Golgi network to the exterior of the cell. Proteins present in regulated secretory vesicles are required to be rapidly exocytosed in large amounts upon a specific stimulus.
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Author Spotlight: Optimizing CFPS Systems for Synthetic Cell Construction
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SecYEG-mediated translocation in a model synthetic cell.

Ludo L J Schoenmakers1, Max J den Uijl2, Jelle L Postma3

  • 1Physical-Organic Chemistry, Institute for Molecules and Materials, Radboud University, Nijmegen 6525AJ, The Netherlands.

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Summary

Researchers successfully incorporated the SecYEG translocase into giant unilamellar vesicles (GUVs), creating SecGUVs. This breakthrough advances synthetic cell development by enabling functional membrane protein integration for growth and division.

Keywords:
Sec translocongiant unilamellar vesiclesliposomessynthetic celltranslocation

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Area of Science:

  • Synthetic biology
  • Biochemistry
  • Membrane protein biophysics

Background:

  • Giant unilamellar vesicles (GUVs) are crucial model systems for synthetic cell research.
  • Incorporating functional membrane proteins into GUVs remains a significant challenge for creating advanced synthetic cells.
  • Key membrane proteins are needed for essential cellular functions like transport, energy conversion, and replication.

Purpose of the Study:

  • To develop a method for incorporating the bacterial translocase SecYEG into GUVs.
  • To create functionalized GUVs (SecGUVs) suitable for large-scale synthetic biology applications.
  • To demonstrate that SecYEG retains its translocation activity within the GUVs.

Main Methods:

  • Reconstitution of the SecYEG protein complex into small unilamellar vesicles (SUVs).
  • Fusion of SecYEG-containing SUVs with pre-formed GUVs to generate SecGUVs.
  • Assessment of SecYEG translocation efficiency within the engineered GUVs.

Main Results:

  • Successfully produced SecGUVs by fusing SecYEG-reconstituted SUVs with GUVs.
  • Maintained a high protein:lipid ratio in the engineered SecGUVs.
  • Demonstrated that SecYEG translocation efficiency is not compromised by its incorporation into GUVs.

Conclusions:

  • The developed method enables the creation of robust, functional proteo-GUVs.
  • SecGUVs are versatile compartments for advancing synthetic cell formation and growth.
  • This work provides a foundation for engineering more complex synthetic cellular systems.