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Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans
Syntaxin-1 is the central SNARE protein for neuronal exocytosis. It interacts with Munc18-1 through its cytoplasmic domains, including the N-terminal peptide (N-peptide). Here we examine the role of the N-peptide binding in two conformational states (“closed” vs. “open”) of syntaxin-1 using PC12 cel...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750926/ https://www.ncbi.nlm.nih.gov/pubmed/26700321 http://dx.doi.org/10.1091/mbc.E15-09-0638 |
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author | Park, Seungmee Bin, Na-Ryum Michael Rajah, Maaran Kim, Byungjin Chou, Ting-Chieh Kang, Soo-young Ann Sugita, Kyoko Parsaud, Leon Smith, Matthew Monnier, Philippe P. Ikura, Mitsuhiko Zhen, Mei Sugita, Shuzo |
author_facet | Park, Seungmee Bin, Na-Ryum Michael Rajah, Maaran Kim, Byungjin Chou, Ting-Chieh Kang, Soo-young Ann Sugita, Kyoko Parsaud, Leon Smith, Matthew Monnier, Philippe P. Ikura, Mitsuhiko Zhen, Mei Sugita, Shuzo |
author_sort | Park, Seungmee |
collection | PubMed |
description | Syntaxin-1 is the central SNARE protein for neuronal exocytosis. It interacts with Munc18-1 through its cytoplasmic domains, including the N-terminal peptide (N-peptide). Here we examine the role of the N-peptide binding in two conformational states (“closed” vs. “open”) of syntaxin-1 using PC12 cells and Caenorhabditis elegans. We show that expression of “closed” syntaxin-1A carrying N-terminal single point mutations (D3R, L8A) that perturb interaction with the hydrophobic pocket of Munc18-1 rescues impaired secretion in syntaxin-1–depleted PC12 cells and the lethality and lethargy of unc-64 (C. elegans orthologue of syntaxin-1)-null mutants. Conversely, expression of the “open” syntaxin-1A harboring the same mutations fails to rescue the impairments. Biochemically, the L8A mutation alone slightly weakens the binding between “closed” syntaxin-1A and Munc18-1, whereas the same mutation in the “open” syntaxin-1A disrupts it. Our results reveal a striking interplay between the syntaxin-1 N-peptide and the conformational state of the protein. We propose that the N-peptide plays a critical role in intracellular trafficking of syntaxin-1, which is dependent on the conformational state of this protein. Surprisingly, however, the N-peptide binding mode seems dispensable for SNARE-mediated exocytosis per se, as long as the protein is trafficked to the plasma membrane. |
format | Online Article Text |
id | pubmed-4750926 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-47509262016-04-30 Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans Park, Seungmee Bin, Na-Ryum Michael Rajah, Maaran Kim, Byungjin Chou, Ting-Chieh Kang, Soo-young Ann Sugita, Kyoko Parsaud, Leon Smith, Matthew Monnier, Philippe P. Ikura, Mitsuhiko Zhen, Mei Sugita, Shuzo Mol Biol Cell Articles Syntaxin-1 is the central SNARE protein for neuronal exocytosis. It interacts with Munc18-1 through its cytoplasmic domains, including the N-terminal peptide (N-peptide). Here we examine the role of the N-peptide binding in two conformational states (“closed” vs. “open”) of syntaxin-1 using PC12 cells and Caenorhabditis elegans. We show that expression of “closed” syntaxin-1A carrying N-terminal single point mutations (D3R, L8A) that perturb interaction with the hydrophobic pocket of Munc18-1 rescues impaired secretion in syntaxin-1–depleted PC12 cells and the lethality and lethargy of unc-64 (C. elegans orthologue of syntaxin-1)-null mutants. Conversely, expression of the “open” syntaxin-1A harboring the same mutations fails to rescue the impairments. Biochemically, the L8A mutation alone slightly weakens the binding between “closed” syntaxin-1A and Munc18-1, whereas the same mutation in the “open” syntaxin-1A disrupts it. Our results reveal a striking interplay between the syntaxin-1 N-peptide and the conformational state of the protein. We propose that the N-peptide plays a critical role in intracellular trafficking of syntaxin-1, which is dependent on the conformational state of this protein. Surprisingly, however, the N-peptide binding mode seems dispensable for SNARE-mediated exocytosis per se, as long as the protein is trafficked to the plasma membrane. The American Society for Cell Biology 2016-02-15 /pmc/articles/PMC4750926/ /pubmed/26700321 http://dx.doi.org/10.1091/mbc.E15-09-0638 Text en © 2016 Park et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Park, Seungmee Bin, Na-Ryum Michael Rajah, Maaran Kim, Byungjin Chou, Ting-Chieh Kang, Soo-young Ann Sugita, Kyoko Parsaud, Leon Smith, Matthew Monnier, Philippe P. Ikura, Mitsuhiko Zhen, Mei Sugita, Shuzo Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans |
title | Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans |
title_full | Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans |
title_fullStr | Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans |
title_full_unstemmed | Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans |
title_short | Conformational states of syntaxin-1 govern the necessity of N-peptide binding in exocytosis of PC12 cells and Caenorhabditis elegans |
title_sort | conformational states of syntaxin-1 govern the necessity of n-peptide binding in exocytosis of pc12 cells and caenorhabditis elegans |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750926/ https://www.ncbi.nlm.nih.gov/pubmed/26700321 http://dx.doi.org/10.1091/mbc.E15-09-0638 |
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