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Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane

The SNAREs SNAP25 and SNAP23 are proteins that are initially cytosolic after translation, but then become stably attached to the cell membrane through palmitoylation of cysteine residues. For palmitoylation to occur, membrane association is a prerequisite, but it is unclear which motif may increase...

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Autores principales: Weber, Pascal, Batoulis, Helena, Rink, Kerstin M, Dahlhoff, Stefan, Pinkwart, Kerstin, Söllner, Thomas H, Lang, Thorsten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5362264/
https://www.ncbi.nlm.nih.gov/pubmed/28240595
http://dx.doi.org/10.7554/eLife.19394
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author Weber, Pascal
Batoulis, Helena
Rink, Kerstin M
Dahlhoff, Stefan
Pinkwart, Kerstin
Söllner, Thomas H
Lang, Thorsten
author_facet Weber, Pascal
Batoulis, Helena
Rink, Kerstin M
Dahlhoff, Stefan
Pinkwart, Kerstin
Söllner, Thomas H
Lang, Thorsten
author_sort Weber, Pascal
collection PubMed
description The SNAREs SNAP25 and SNAP23 are proteins that are initially cytosolic after translation, but then become stably attached to the cell membrane through palmitoylation of cysteine residues. For palmitoylation to occur, membrane association is a prerequisite, but it is unclear which motif may increase the affinities of the proteins for the target membrane. In experiments with rat neuroendocrine cells, we find that a few basic amino acids in the cysteine-rich region of SNAP25 and SNAP23 are essential for plasma membrane targeting. Reconstitution of membrane-protein binding in a liposome assay shows that the mechanism involves protein electrostatics between basic amino acid residues and acidic lipids such as phosphoinositides that play a primary role in these interactions. Hence, we identify an electrostatic anchoring mechanism underlying initial plasma membrane contact by SNARE proteins, which subsequently become palmitoylated at the plasma membrane. DOI: http://dx.doi.org/10.7554/eLife.19394.001
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spelling pubmed-53622642017-03-24 Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane Weber, Pascal Batoulis, Helena Rink, Kerstin M Dahlhoff, Stefan Pinkwart, Kerstin Söllner, Thomas H Lang, Thorsten eLife Biophysics and Structural Biology The SNAREs SNAP25 and SNAP23 are proteins that are initially cytosolic after translation, but then become stably attached to the cell membrane through palmitoylation of cysteine residues. For palmitoylation to occur, membrane association is a prerequisite, but it is unclear which motif may increase the affinities of the proteins for the target membrane. In experiments with rat neuroendocrine cells, we find that a few basic amino acids in the cysteine-rich region of SNAP25 and SNAP23 are essential for plasma membrane targeting. Reconstitution of membrane-protein binding in a liposome assay shows that the mechanism involves protein electrostatics between basic amino acid residues and acidic lipids such as phosphoinositides that play a primary role in these interactions. Hence, we identify an electrostatic anchoring mechanism underlying initial plasma membrane contact by SNARE proteins, which subsequently become palmitoylated at the plasma membrane. DOI: http://dx.doi.org/10.7554/eLife.19394.001 eLife Sciences Publications, Ltd 2017-02-27 /pmc/articles/PMC5362264/ /pubmed/28240595 http://dx.doi.org/10.7554/eLife.19394 Text en © 2017, Weber et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biophysics and Structural Biology
Weber, Pascal
Batoulis, Helena
Rink, Kerstin M
Dahlhoff, Stefan
Pinkwart, Kerstin
Söllner, Thomas H
Lang, Thorsten
Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane
title Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane
title_full Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane
title_fullStr Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane
title_full_unstemmed Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane
title_short Electrostatic anchoring precedes stable membrane attachment of SNAP25/SNAP23 to the plasma membrane
title_sort electrostatic anchoring precedes stable membrane attachment of snap25/snap23 to the plasma membrane
topic Biophysics and Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5362264/
https://www.ncbi.nlm.nih.gov/pubmed/28240595
http://dx.doi.org/10.7554/eLife.19394
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