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Structural requirements for steady-state localization of the vesicular acetylcholine transporter.
- Source :
-
Journal of neurochemistry [J Neurochem] 2005 Aug; Vol. 94 (4), pp. 957-69. - Publication Year :
- 2005
-
Abstract
- The vesicular acetylcholine transporter (VAChT) regulates the amount of acetylcholine stored in synaptic vesicles. However, the mechanisms that control the targeting of VAChT and other synaptic vesicle proteins are still poorly comprehended. These processes are likely to depend, at least partially, on structural determinants present in the primary sequence of the protein. Here, we use site-directed mutagenesis to evaluate the contribution of the C-terminal tail of VAChT to the targeting of this transporter to synaptic-like microvesicles in cholinergic SN56 cells. We found that residues 481-490 contain the trafficking information necessary for VAChT localization and that within this region L485 and L486 are strictly necessary. Deletion and alanine-scanning mutants lacking most of the carboxyl tail of VAChT, but containing residues 481-490, were still targeted to microvesicles. Moreover, we found that clathrin-mediated endocytosis of VAChT is required for targeting to microvesicles in SN56 and PC12 cells. The data provide novel information on the mechanisms and structural determinants necessary for VAChT localization to synaptic vesicles.
- Subjects :
- Amino Acid Sequence
Animals
Cell Line
Clathrin physiology
Endocytosis physiology
Mice
Molecular Sequence Data
Mutagenesis, Site-Directed
PC12 Cells
Protein Conformation
Rats
Tissue Distribution
Vesicular Acetylcholine Transport Proteins
Homeostasis
Membrane Transport Proteins genetics
Membrane Transport Proteins metabolism
Synaptic Vesicles metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0022-3042
- Volume :
- 94
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Journal of neurochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 16092939
- Full Text :
- https://doi.org/10.1111/j.1471-4159.2005.03244.x