Stepwise assembly of functionally active transport vesicles

Cell. 1993 Dec 3;75(5):1015-25. doi: 10.1016/0092-8674(93)90545-2.

Abstract

Budding of COP-coated vesicles (the likely carriers of newly synthesized proteins from the endoplasmic reticulum through the Golgi stack) from Golgi cisternae requires ADP-ribosylation factor (ARF), coatomer proteins from the cytosol, GTP, and fatty acyl-coenzyme A (CoA). The assembly of coated buds on the membranes requires coatomer, ARF, and GTP. When palmitoyl-CoA is added, membrane fission occurs at the coated bud, releasing coated vesicles. We show that COP-coated vesicles can be generated stepwise in vitro and isolated in a functionally active form, demonstrating that the minimal set of cytosolic components required for their formation as well as principal steps in their assembly have been identified.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ADP-Ribosylation Factors
  • Acyl Coenzyme A / metabolism
  • Animals
  • Biological Transport, Active
  • CHO Cells
  • Cell Compartmentation
  • Coatomer Protein
  • Cricetinae
  • GTP-Binding Proteins / metabolism*
  • Golgi Apparatus / metabolism*
  • Guanosine Triphosphate / metabolism
  • In Vitro Techniques
  • Intracellular Membranes / metabolism*
  • Membrane Glycoproteins / metabolism
  • Membrane Proteins / metabolism*
  • Microtubule-Associated Proteins / metabolism*
  • Morphogenesis
  • Organelles / metabolism
  • Organelles / ultrastructure*

Substances

  • Acyl Coenzyme A
  • Coatomer Protein
  • Membrane Glycoproteins
  • Membrane Proteins
  • Microtubule-Associated Proteins
  • Guanosine Triphosphate
  • GTP-Binding Proteins
  • ADP-Ribosylation Factors