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PMID: 16195346 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Alternative splicing of SNAP-25 regulates secretion through nonconservative substitutions in the SNARE domain.

Molecular biology of the cell ·Vol. 16 ·No. 12 ·2005-12-00 ·Pages 5675-85

Nagy G, Milosevic I, Fasshauer D, Müller EM, de Groot BL, Lang T, Wilson MC, Sørensen JB

Abstract

The essential membrane fusion apparatus in mammalian cells, the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, consists of four alpha-helices formed by three proteins: SNAP-25, syntaxin 1, and synaptobrevin 2. SNAP-25 contributes two helices to the complex and is targeted to the plasma membrane by palmitoylation of four cysteines in the linker region. It is alternatively spliced into two forms, SNAP-25a and SNAP-25b, differing by nine amino acids substitutions. When expressed in chromaffin cells from SNAP-25 null mice, the isoforms support different levels of secretion. Here, we investigated the basis of that different secretory phenotype. We found that two nonconservative substitutions in the N-terminal SNARE domain and not the different localization of one palmitoylated cysteine cause the functional difference between the isoforms. Biochemical and molecular dynamic simulation experiments revealed that the two substitutions do not regulate secretion by affecting the property of SNARE complex itself, but rather make the SNAP-25b-containing SNARE complex more available for the interaction with accessory factor(s).

MeSH Terms
Alternative Splicing Amino Acid Sequence Amino Acid Substitution Animals Chromaffin Cells/physiology Circular Dichroism Computer Simulation Exons/genetics Mice Mice, Knockout Molecular Sequence Data SNARE Proteins/chemistry,genetics Synaptosomal-Associated Protein 25/chemistry,deficiency,genetics
Chemicals
SNARE Proteins Synaptosomal-Associated Protein 25
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Nagy Gábor
Department of Membrane Biophysics, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
Milosevic Ira
Fasshauer Dirk
Müller E Matthias
de Groot Bert L
Lang Thorsten
Wilson Michael C
Sørensen Jakob B
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-07-0595
Published
2005-12-00
Epub
2005-00-29
Pages
5675-85
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1289412
Subset
IM
Grants
NIMH NIH HHS · R01 MH048989 · United States
NIMH NIH HHS · MH-48989 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-07-0595 PMC Full Text View journal details
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