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Vol. 9, Issue 6, 1437-1448, June 1998


and
*Centre National de la Recherche Scientifique Unité
Mixte de Recherche 144 "Compartimentation et Dynamique
Cellulaires," Institut Curie, F-75248 Paris Cedex 05, France;
The importance of soluble N-ethyl
maleimide (NEM)-sensitive fusion protein (NSF) attachment protein
(SNAP) receptors (SNAREs) in synaptic vesicle exocytosis is well
established because it has been demonstrated that clostridial
neurotoxins (NTs) proteolyze the vesicle SNAREs (v-SNAREs)
vesicle-associated membrane protein (VAMP)/brevins and their partners,
the target SNAREs (t-SNAREs) syntaxin 1 and SNAP25. Yet, several
exocytotic events, including apical exocytosis in epithelial cells, are
insensitive to numerous clostridial NTs, suggesting the presence of
SNARE-independent mechanisms of exocytosis. In this study we found that
syntaxin 3, SNAP23, and a newly identified VAMP/brevin, tetanus
neurotoxin (TeNT)-insensitive VAMP (TI-VAMP), are insensitive to
clostridial NTs. In epithelial cells, TI-VAMP-containing vesicles were
concentrated in the apical domain, and the protein was detected at the
apical plasma membrane by immunogold labeling on ultrathin
cryosections. Syntaxin 3 and SNAP23 were codistributed at the apical
plasma membrane where they formed NEM-dependent SNARE complexes with TI-VAMP and cellubrevin. We suggest that TI-VAMP, SNAP23, and syntaxin
3 can participate in exocytotic processes at the apical plasma membrane
of epithelial cells and, more generally, domain-specific exocytosis in
clostridial NT-resistant pathways.
Medizinische Hochschule Hannover, Department of
Biochemistry, D-30623 Hannover, Germany; and
§Department
of Pharmacology, University of California at San Diego, La Jolla,
California 92093-0636
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