Cargando…
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...
Autores principales: | , , , , , , |
---|---|
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 |
_version_ | 1782516931822092288 |
---|---|
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 |
format | Online Article Text |
id | pubmed-5362264 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT weberpascal electrostaticanchoringprecedesstablemembraneattachmentofsnap25snap23totheplasmamembrane AT batoulishelena electrostaticanchoringprecedesstablemembraneattachmentofsnap25snap23totheplasmamembrane AT rinkkerstinm electrostaticanchoringprecedesstablemembraneattachmentofsnap25snap23totheplasmamembrane AT dahlhoffstefan electrostaticanchoringprecedesstablemembraneattachmentofsnap25snap23totheplasmamembrane AT pinkwartkerstin electrostaticanchoringprecedesstablemembraneattachmentofsnap25snap23totheplasmamembrane AT sollnerthomash electrostaticanchoringprecedesstablemembraneattachmentofsnap25snap23totheplasmamembrane AT langthorsten electrostaticanchoringprecedesstablemembraneattachmentofsnap25snap23totheplasmamembrane |