title: A novel class of clathrin-coated vesicles budding from endosomes.
abstract: Clathrin-coated vesicles transport selective integral membrane proteins from the plasma membrane to endosomes and from the TGN to endosomes. Recycling of proteins from endosomes to the plasma membrane occurs via unidentified vesicles. To study this pathway, we used a novel technique that allows for the immunoelectron microscopic examination of transferrin receptor-containing endosomes in nonsectioned cells. Endosomes were identified as separate discontinuous tubular-vesicular entities. Each endosome was decorated, mainly on the tubules, with many clathrin-coated buds. Endosome-associated clathrin-coated buds were discerned from plasma membrane-derived clathrin-coated vesicles by three criteria: size (60 nm and 100 nm, respectively), continuity with endosomes, and the lack of labeling for alpha-adaptin. They were also distinguished from TGN-derived clathrin-coated vesicles by their location at the periphery of the cell, size, and the lack of labeling for gamma-adaptin. In the presence of brefeldin A, a large continuous endosomal network was formed. Transferrin receptor recycling as well as the formation of clathrin-coated pits at endosomes was inhibited in the presence of brefeldin A. Together with the localization of transferrin receptors at endosome-associated buds, this indicates that a novel class of clathrin-coated vesicles serves an exit pathway from endosomes. The target organelles for endosome-derived clathrin-coated vesicles remain, however, to be identified.
authors.affiliation: Department of Cell Biology, Faculty of Medicine, Utrecht Universiteit, The Netherlands.
authors.forename: W
authors.initials: W
authors.lastname: Stoorvogel
authors.forename: V
authors.initials: V
authors.lastname: Oorschot
authors.forename: H J
authors.initials: HJ
authors.lastname: Geuze
mesh_terms: D033982:Adaptor Protein Complex gamma Subunits; D001692:Biological Transport; D002462:Cell Membrane / Q000502:physiology; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002966:Clathrin / Q000302:isolation & purification*; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000145:classification* / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000502:physiology; D006651:Histocytochemistry; D006735:Horseradish Peroxidase; D007425:Intracellular Membranes / Q000145:classification* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000302:isolation & purification; D011990:Receptors, Transferrin / Q000302:isolation & purification*
chemical_list: D033982:Adaptor Protein Complex gamma Subunits; D002966:Clathrin; D008565:Membrane Proteins; D011990:Receptors, Transferrin; D006735:Horseradish Peroxidase