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Vol. 11, Issue 10, 3289-3298, October 2000
Endobrevin/VAMP-8 is an R-SNARE localized to endosomes, but
it is unknown in which intracellular fusion step it operates. Using
subcellular fractionation and quantitative immunogold electron microscopy, we found that endobrevin/VAMP-8 is present on all membranes
known to communicate with early endosomes, including the plasma
membrane, clathrin-coated pits, late endosomes, and membranes of the
trans-Golgi network. Affinity-purified antibodies that
block the ability of endobrevin/VAMP-8 to form SNARE core complexes
potently inhibit homotypic fusion of both early and late endosomes in
vitro. Fab fragments were as active as intact immunoglobulin Gs.
Recombinant endobrevin/VAMP-8 inhibited both fusion reactions with
similar potency. We conclude that endobrevin/VAMP-8 operates as an
R-SNARE in the homotypic fusion of early and late endosomes.
Department of Neurobiology, Max-Planck-Institute for
Biophysical Chemistry, Göttingen, Germany;
Institute for Cell Biology, University of Bonn, Bonn,
Germany; and §Institute for Biochemistry II, University of
Göttingen, Göttingen, Germany
Corresponding author. E-mail address:
rjahn{at}gwdg.de.
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