Intracellular site of asialoglycoprotein receptor-ligand uncoupling: double-label immunoelectron microscopy during receptor-mediated endocytosis

Cell. 1983 Jan;32(1):277-87. doi: 10.1016/0092-8674(83)90518-4.

Abstract

In rats infused with asialoglycoprotein for 60 min, receptor-mediated endocytosis of the ligand occurred exclusively in hepatic parenchymal cells. We have used double-label immunoelectron microscopy on ultrathin cryosections of rat liver to identify the site at which the asialoglycoprotein receptor and its ligand dissociate following their common endocytosis. Asialoglycoprotein receptor, ligand and clathrin were identified and quantitated by the use of monospecific antibodies followed by gold-protein A complexes. Both receptor and ligand were found associated with the membrane of clathrin-coated vesicles close to the cell surface. We identified other vesicles that contained ligand accumulated within the lumen. The membranes of these latter vesicles contained little receptor, but receptor was concentrated in tubular extensions that were largely free of ligand. We call this organelle CURL (compartment of uncoupling of receptor and ligand). CURL vesicles appear to transform into secondary lysosomes, wherein the ligand is degraded. The tubular vesicles are, we propose, an intermediate in recycling the receptor to the cell surface.

Publication types

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

MeSH terms

  • Animals
  • Asialoglycoprotein Receptor
  • Cell Compartmentation
  • Coated Pits, Cell-Membrane / metabolism
  • Endocytosis*
  • Endosomes / metabolism
  • Gold
  • Immunologic Techniques
  • Intracellular Membranes / metabolism*
  • Liver / metabolism*
  • Microscopy, Electron
  • Rats
  • Receptors, Cell Surface / metabolism*

Substances

  • Asialoglycoprotein Receptor
  • Receptors, Cell Surface
  • Gold