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J. Biol. Chem., Vol. 277, Issue 38, 35357-35363, September 20, 2002
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From the Secretory carrier membrane proteins (SCAMPs) are
conserved four transmembrane-spanning proteins associated with
recycling vesicular carriers. In mast cells, as in other cell types,
SCAMPs 1 and 2 are present in secretory granule membranes and other
intracellular membranes. We now demonstrate a population of these
SCAMPs in plasma membranes. Although small, this population partially
colocalizes with SNARE proteins SNAP-23 and syntaxin 4. A fraction of
SCAMPs 1 and 2 also coimmunoprecipitates with SNAP-23. An oligopeptide, E peptide, within the cytoplasmic segment linking the second and third
transmembrane spans, particularly of SCAMP2, potently inhibits exocytosis in streptolysin O-permeabilized mast cells. The E peptide is
unique to SCAMPs and highly conserved among SCAMP isoforms, and minor
changes in its sequence abrogate inhibition. It blocks fusion beyond
the putative docking step where granules contact the cell surface and
each other during compound exocytosis. Blockade is also beyond
Ca2+/ATP-dependent relocation of SNAP-23,
which regulates compound exocytosis, and beyond
ATP-dependent priming of fusion. Kinetic ordering of
exocytotic inhibitors has shown that E peptide acts later than other
perturbants at a stage closely associated with membrane fusion. These
findings identify a new reagent for analyzing the final stage of
exocytosis and point to the likely action of SCAMP2 in this process.
Perturbation of a Very Late Step of Regulated Exocytosis by a
Secretory Carrier Membrane Protein (SCAMP2)-derived Peptide*
,,¶
Department of Cell Biology, University of
Virginia Health Sciences Center and the § Department of
Chemistry, University of Virginia,
Charlottesville, Virginia 22908
*
This work was supported by National Institutes of Health
Grants DE09655 and AI47150.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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