Purification of a novel class of coated vesicles mediating biosynthetic protein transport through the Golgi stack.
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.
Malhotra
V
V
Department of Biology Princeton University, New Jersey 08544.
Serafini
T
T
Orci
L
L
Shepherd
J C
JC
Rothman
J E
JE
D000818:Animals; D001692:Biological Transport; D002458:Cell Fractionation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D006056:Golgi Apparatus / Q000378:metabolism / Q000648:ultrastructure*; D007425:Intracellular Membranes / Q000032:analysis / Q000502:physiology / Q000648:ultrastructure*; D008099:Liver / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000378:metabolism / Q000493:pharmacokinetics*; D011817:Rabbits
D010455:Peptides; D011506:Proteins