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The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix
Two-pore channels (TPCs) constitute a family of intracellular cation channels with diverse permeation properties and functions in animals and plants. In the model plant Arabidopsis, the vacuolar cation channel TPC1 is involved in propagation of calcium waves and in cation homeostasis. Here, we disco...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4894940/ https://www.ncbi.nlm.nih.gov/pubmed/26781468 http://dx.doi.org/10.1007/s00018-016-2131-3 |
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author | Larisch, Nina Kirsch, Sonja A. Schambony, Alexandra Studtrucker, Tanja Böckmann, Rainer A. Dietrich, Petra |
author_facet | Larisch, Nina Kirsch, Sonja A. Schambony, Alexandra Studtrucker, Tanja Böckmann, Rainer A. Dietrich, Petra |
author_sort | Larisch, Nina |
collection | PubMed |
description | Two-pore channels (TPCs) constitute a family of intracellular cation channels with diverse permeation properties and functions in animals and plants. In the model plant Arabidopsis, the vacuolar cation channel TPC1 is involved in propagation of calcium waves and in cation homeostasis. Here, we discovered that the dimerization of a predicted helix within the carboxyl-terminus (CTH) is essential for the activity of TPC1. Bimolecular fluorescence complementation and co-immunoprecipitation demonstrated the interaction of the two C-termini and pointed towards the involvement of the CTH in this process. Synthetic CTH peptides dimerized with a dissociation constant of 3.9 µM. Disruption of this domain in TPC1 either by deletion or point mutations impeded the dimerization and cation transport. The homo-dimerization of the CTH was analyzed in silico using coarse-grained molecular dynamics (MD) simulations for the study of aggregation, followed by atomistic MD simulations. The simulations revealed that the helical region of the wild type, but not a mutated CTH forms a highly stable, antiparallel dimer with characteristics of a coiled-coil. We propose that the voltage- and Ca(2+)-sensitive conformation of TPC1 depends on C-terminal dimerization, adding an additional layer to the complex regulation of two-pore cation channels. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00018-016-2131-3) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4894940 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-48949402016-06-20 The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix Larisch, Nina Kirsch, Sonja A. Schambony, Alexandra Studtrucker, Tanja Böckmann, Rainer A. Dietrich, Petra Cell Mol Life Sci Original Article Two-pore channels (TPCs) constitute a family of intracellular cation channels with diverse permeation properties and functions in animals and plants. In the model plant Arabidopsis, the vacuolar cation channel TPC1 is involved in propagation of calcium waves and in cation homeostasis. Here, we discovered that the dimerization of a predicted helix within the carboxyl-terminus (CTH) is essential for the activity of TPC1. Bimolecular fluorescence complementation and co-immunoprecipitation demonstrated the interaction of the two C-termini and pointed towards the involvement of the CTH in this process. Synthetic CTH peptides dimerized with a dissociation constant of 3.9 µM. Disruption of this domain in TPC1 either by deletion or point mutations impeded the dimerization and cation transport. The homo-dimerization of the CTH was analyzed in silico using coarse-grained molecular dynamics (MD) simulations for the study of aggregation, followed by atomistic MD simulations. The simulations revealed that the helical region of the wild type, but not a mutated CTH forms a highly stable, antiparallel dimer with characteristics of a coiled-coil. We propose that the voltage- and Ca(2+)-sensitive conformation of TPC1 depends on C-terminal dimerization, adding an additional layer to the complex regulation of two-pore cation channels. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00018-016-2131-3) contains supplementary material, which is available to authorized users. Springer International Publishing 2016-01-18 2016 /pmc/articles/PMC4894940/ /pubmed/26781468 http://dx.doi.org/10.1007/s00018-016-2131-3 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Article Larisch, Nina Kirsch, Sonja A. Schambony, Alexandra Studtrucker, Tanja Böckmann, Rainer A. Dietrich, Petra The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix |
title | The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix |
title_full | The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix |
title_fullStr | The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix |
title_full_unstemmed | The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix |
title_short | The function of the two-pore channel TPC1 depends on dimerization of its carboxy-terminal helix |
title_sort | function of the two-pore channel tpc1 depends on dimerization of its carboxy-terminal helix |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4894940/ https://www.ncbi.nlm.nih.gov/pubmed/26781468 http://dx.doi.org/10.1007/s00018-016-2131-3 |
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