title: Purification of a novel class of coated vesicles mediating biosynthetic protein transport through the Golgi stack.
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.
authors.affiliation: Department of Biology Princeton University, New Jersey 08544.
authors.forename: V
authors.initials: V
authors.lastname: Malhotra
authors.forename: T
authors.initials: T
authors.lastname: Serafini
authors.forename: L
authors.initials: L
authors.lastname: Orci
authors.forename: J C
authors.initials: JC
authors.lastname: Shepherd
authors.forename: J E
authors.initials: JE
authors.lastname: Rothman
mesh_terms: 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
chemical_list: D010455:Peptides; D011506:Proteins