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Related Concept Videos

Pinching-off of Coated Vesicles01:32

Pinching-off of Coated Vesicles

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Vesicle budding is orchestrated by distinct cytosolic proteins such as adaptor proteins, coat proteins, and GTPases. To initiate vesicle budding, membrane-bending proteins containing crescent-shaped BAR domains bind to the lipid heads in the bilayer and distort the membrane to form a protein-coated vesicle bud. Adaptors proteins such as AP2 for clathrin-coated vesicles can nucleate on the deformed membrane. Finally, coat proteins such as clathrin or COPI and COPII assemble into a coat forming...
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Membrane-enclosed structures called vesicles transport proteins and lipids across the cell. The vesicles derive their cargo from the plasma membrane, Golgi, ER, or endosome. Coated vesicles are spherical, protein-coated carriers with a 50–100 nm diameter that mediate bidirectional transport between the ER and the Golgi. The distribution of proteins between the ER and Golgi complex is dynamic and is maintained by different coated vesicles. Their formation is driven by the assembly of...
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After budding out from the ER membrane, some COPII vesicles lose their coat and fuse with one another to form larger vesicles and interconnected tubules called vesicular tubular clusters or VTCs. These clusters constitute a compartment at the ER-Golgi interface known as ERGIC (Endoplasmic Reticulum Golgi Intermediate Compartment). The ERGIC is a mobile membrane-bound cargo transport system that sorts proteins secreted from ER and delivers them to the Golgi.
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Related Experiment Video

Updated: Jun 17, 2025

In Vitro Reconstitution of the Actin Cytoskeleton Inside Giant Unilamellar Vesicles
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In Vitro Reconstitution of Plant COPII Vesicles.

Baiying Li1, Liwen Jiang2

  • 1Department of Biology, Hong Kong Baptist University, Hong Kong, China. baiying@hkbu.edu.hk.

Methods in Molecular Biology (Clifton, N.J.)
|August 8, 2024
PubMed
Summary

This study details the preparation of plant microsomes and cytosol for in vitro reconstitution of COPII vesicles. This method aids in studying endoplasmic reticulum-to-Golgi transport in plants.

Keywords:
Endomembrane traffickingIn vitro reconstitutionPlant COPII vesicle

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Area of Science:

  • Plant cell biology
  • Molecular and Cellular Biology
  • Biochemistry

Background:

  • In vitro reconstitution studies are crucial for dissecting complex biochemical processes like vesicle transport.
  • COPII vesicle reconstitution in yeast and mammals has significantly advanced understanding of the endoplasmic reticulum-to-Golgi pathway.
  • Cell-free systems allow for controlled investigation of vesicle formation and regulation.

Purpose of the Study:

  • To describe the preparation of plant microsomes and cytosol for the in vitro reconstitution of plant COPII vesicles.
  • To enable detailed biochemical and microscopical analyses of plant COPII vesicle formation and function.

Main Methods:

  • Preparation of plant microsomes.
  • Preparation of plant cytosol.
  • In vitro reconstitution of plant COPII vesicles using prepared microsomes and cytosol.

Main Results:

  • Successful preparation of plant microsomes and cytosol suitable for COPII vesicle reconstitution.
  • The described method yields purified plant COPII vesicles for subsequent analyses.

Conclusions:

  • The described protocol facilitates the in vitro reconstitution of plant COPII vesicles.
  • This system provides a valuable tool for studying plant-specific anterograde trafficking mechanisms.
  • Purified vesicles can be utilized for further biochemical and microscopical investigations.