Purification of a novel class of coated vesicles mediating biosynthetic protein transport through the Golgi stack

Cell. 1989 Jul 28;58(2):329-36. doi: 10.1016/0092-8674(89)90847-7.

Abstract

We describe a scheme for the purification of the nonclathrin-coated vesicles that mediate transport of proteins between Golgi cisternae and probably from ER to Golgi. These "Golgi-derived coated vesicles" accumulate when Golgi membranes are incubated with ATP and cytosol in the presence of GTP gamma S, a compound that blocks vesicle fusion. The coated vesicles dissociate from the Golgi cisternae in high salt and can then be purified by employing differential and density gradient centrifugation. Golgi-derived coated vesicles have a putative polypeptide composition that is distinct from both cytosol and Golgi membranes, as well as from that of clathrin-coated vesicles.

Publication types

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

MeSH terms

  • Animals
  • Biological Transport
  • Cell Fractionation / methods*
  • Centrifugation, Density Gradient
  • Electrophoresis, Polyacrylamide Gel
  • Golgi Apparatus / metabolism
  • Golgi Apparatus / ultrastructure*
  • Intracellular Membranes / analysis
  • Intracellular Membranes / physiology
  • Intracellular Membranes / ultrastructure*
  • Liver / cytology
  • Liver / metabolism
  • Liver / ultrastructure
  • Peptides / analysis
  • Proteins / metabolism
  • Proteins / pharmacokinetics*
  • Rabbits

Substances

  • Peptides
  • Proteins