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J. Biol. Chem., Vol. 263, Issue 33, 17213-17216, 11, 1988
LS Mayorga, R Diaz and PD Stahl
Receptor-mediated endocytosis involves the transport of receptor-ligand
complexes from the cell surface to an intracellular endocytic compartment.
This study shows that plasma membrane-derived vesicles containing
receptor-bound ligands (e.g. aggregated anti-dinitrophenol (DNP) IgG bound
to Fc receptors) fuse with early endosomes containing
DNP-beta-glucuronidase in a cell-free system. Plasma membrane vesicles were
generated by homogenization of cells that had been allowed to bind ligands
at 4 degrees C. Fusion between vesicles containing the two probes was
assessed by (i) the formation of anti-DNP IgG-DNP-beta- glucuronidase
complexes and (ii) the colocalization within closed vesicles of two
different sizes of colloidal gold coated with ligands. Fusion required ATP,
cytosol, and KCl. The requirements were similar to those described for
endosome-endosome fusion in in vitro systems. Mild trypsinization of
vesicles prior to their addition to the assay inhibited fusion. When
DNP-beta-glucuronidase was chased into more mature endocytic compartments,
fusion was not observed. The results indicate that cell surface regions
involved in receptor-mediated endocytosis are capable of fusing to early
endosomes. This fusion event may constitute the first step in the transport
of ligands to an intracellular endocytic compartment.
Plasma membrane-derived vesicles containing receptor-ligand complexes are fusogenic with early endosomes in a cell-free system
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110.
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