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Related Experiment Videos

A function for the AP3 coat complex in synaptic vesicle formation from endosomes

V Faúndez1, J T Horng, R B Kelly

  • 1Department of Biochemistry and Biophysics and Hormone Research Institute, University of California, San Francisco 94143-0534, USA.

Cell
|May 20, 1998
PubMed
Summary
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Adaptor protein 3 (AP3) coats synaptic vesicles in vitro, a process dependent on ARF, ATP, and temperature. This finding suggests AP3 is crucial for small vesicle formation from endosomes.

Area of Science:

  • Cell biology
  • Molecular neuroscience
  • Protein complex research

Background:

  • Synaptic vesicle coating is essential for neurotransmission.
  • The role of specific protein complexes in vesicle formation is under investigation.
  • Adaptor protein 3 (AP3) is known to mediate Golgi-to-vacuole transport in yeast.

Purpose of the Study:

  • To investigate the role of adaptor protein 3 (AP3) in synaptic vesicle coating and formation.
  • To determine the requirements for in vitro synaptic vesicle coating.

Main Methods:

  • In vitro coating reactions using brain cytosol and PC12 endosomes.
  • Depletion of AP3 from cytosol.
  • Reconstitution experiments with purified AP3 and recombinant ARF1.

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Main Results:

  • Synaptic vesicle coating in vitro is dependent on ARF, ATP, and temperature.
  • AP3 is the major component of the synaptic vesicle coat.
  • Depletion of AP3 inhibits small vesicle formation from endosomes.
  • Purified AP3 and ARF1 can reconstitute vesicle budding from membranes.

Conclusions:

  • AP3 coating is involved in a pathway of small vesicle formation from endosomes.
  • AP3 plays a significant role in synaptic vesicle biogenesis.