The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes

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 endos...

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Veröffentlicht in:Molecular biology of the cell 2000-10, Vol.11 (10), p.3289-3298
Hauptverfasser: Antonin, W, Holroyd, C, Tikkanen, R, Höning, S, Jahn, R
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Sprache:eng
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Zusammenfassung: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.
ISSN:1059-1524
1939-4586
DOI:10.1091/mbc.11.10.3289