Cargando…
Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding
Transient receptor potential (TRP) cation channels, which are conserved across mammals, flies, fish, sea squirts, worms, and fungi, essentially contribute to cellular Ca(2+) signaling. The activity of the unique TRP channel in yeast, TRP yeast channel 1 (TRPY1), relies on the vacuolar and cytoplasmi...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449268/ https://www.ncbi.nlm.nih.gov/pubmed/34461097 http://dx.doi.org/10.1016/j.jbc.2021.101126 |
_version_ | 1784569396549648384 |
---|---|
author | Amini, Mahnaz Chang, Yiming Wissenbach, Ulrich Flockerzi, Veit Schlenstedt, Gabriel Beck, Andreas |
author_facet | Amini, Mahnaz Chang, Yiming Wissenbach, Ulrich Flockerzi, Veit Schlenstedt, Gabriel Beck, Andreas |
author_sort | Amini, Mahnaz |
collection | PubMed |
description | Transient receptor potential (TRP) cation channels, which are conserved across mammals, flies, fish, sea squirts, worms, and fungi, essentially contribute to cellular Ca(2+) signaling. The activity of the unique TRP channel in yeast, TRP yeast channel 1 (TRPY1), relies on the vacuolar and cytoplasmic Ca(2+) concentration. However, the mechanism(s) of Ca(2+)-dependent regulation of TRPY1 and possible contribution(s) of Ca(2+)-binding proteins are yet not well understood. Our results demonstrate a Ca(2+)-dependent binding of yeast calmodulin (CaM) to TRPY1. TRPY1 activity was increased in the cmd1–6 yeast strain, carrying a non–Ca(2+)-binding CaM mutant, compared with the parent strain expressing wt CaM (Cmd1). Expression of Cmd1 in cmd1–6 yeast rescued the wt phenotype. In addition, in human embryonic kidney 293 cells, hypertonic shock-induced TRPY1-dependent Ca(2+) influx and Ca(2+) release were increased by the CaM antagonist ophiobolin A. We found that coexpression of mammalian CaM impeded the activity of TRPY1 by reinforcing effects of endogenous CaM. Finally, inhibition of TRPY1 by Ca(2+)–CaM required the cytoplasmic amino acid stretch E(33)–Y(92). In summary, our results show that TRPY1 is under inhibitory control of Ca(2+)–CaM and that mammalian CaM can replace yeast CaM for this inhibition. These findings add TRPY1 to the innumerable cellular proteins, which include a variety of ion channels, that use CaM as a constitutive or dissociable Ca(2+)-sensing subunit, and contribute to a better understanding of the modulatory mechanisms of Ca(2+)–CaM. |
format | Online Article Text |
id | pubmed-8449268 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-84492682021-09-24 Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding Amini, Mahnaz Chang, Yiming Wissenbach, Ulrich Flockerzi, Veit Schlenstedt, Gabriel Beck, Andreas J Biol Chem Research Article Transient receptor potential (TRP) cation channels, which are conserved across mammals, flies, fish, sea squirts, worms, and fungi, essentially contribute to cellular Ca(2+) signaling. The activity of the unique TRP channel in yeast, TRP yeast channel 1 (TRPY1), relies on the vacuolar and cytoplasmic Ca(2+) concentration. However, the mechanism(s) of Ca(2+)-dependent regulation of TRPY1 and possible contribution(s) of Ca(2+)-binding proteins are yet not well understood. Our results demonstrate a Ca(2+)-dependent binding of yeast calmodulin (CaM) to TRPY1. TRPY1 activity was increased in the cmd1–6 yeast strain, carrying a non–Ca(2+)-binding CaM mutant, compared with the parent strain expressing wt CaM (Cmd1). Expression of Cmd1 in cmd1–6 yeast rescued the wt phenotype. In addition, in human embryonic kidney 293 cells, hypertonic shock-induced TRPY1-dependent Ca(2+) influx and Ca(2+) release were increased by the CaM antagonist ophiobolin A. We found that coexpression of mammalian CaM impeded the activity of TRPY1 by reinforcing effects of endogenous CaM. Finally, inhibition of TRPY1 by Ca(2+)–CaM required the cytoplasmic amino acid stretch E(33)–Y(92). In summary, our results show that TRPY1 is under inhibitory control of Ca(2+)–CaM and that mammalian CaM can replace yeast CaM for this inhibition. These findings add TRPY1 to the innumerable cellular proteins, which include a variety of ion channels, that use CaM as a constitutive or dissociable Ca(2+)-sensing subunit, and contribute to a better understanding of the modulatory mechanisms of Ca(2+)–CaM. American Society for Biochemistry and Molecular Biology 2021-08-28 /pmc/articles/PMC8449268/ /pubmed/34461097 http://dx.doi.org/10.1016/j.jbc.2021.101126 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Amini, Mahnaz Chang, Yiming Wissenbach, Ulrich Flockerzi, Veit Schlenstedt, Gabriel Beck, Andreas Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding |
title | Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding |
title_full | Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding |
title_fullStr | Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding |
title_full_unstemmed | Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding |
title_short | Activity of the yeast vacuolar TRP channel TRPY1 is inhibited by Ca(2+)–calmodulin binding |
title_sort | activity of the yeast vacuolar trp channel trpy1 is inhibited by ca(2+)–calmodulin binding |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449268/ https://www.ncbi.nlm.nih.gov/pubmed/34461097 http://dx.doi.org/10.1016/j.jbc.2021.101126 |
work_keys_str_mv | AT aminimahnaz activityoftheyeastvacuolartrpchanneltrpy1isinhibitedbyca2calmodulinbinding AT changyiming activityoftheyeastvacuolartrpchanneltrpy1isinhibitedbyca2calmodulinbinding AT wissenbachulrich activityoftheyeastvacuolartrpchanneltrpy1isinhibitedbyca2calmodulinbinding AT flockerziveit activityoftheyeastvacuolartrpchanneltrpy1isinhibitedbyca2calmodulinbinding AT schlenstedtgabriel activityoftheyeastvacuolartrpchanneltrpy1isinhibitedbyca2calmodulinbinding AT beckandreas activityoftheyeastvacuolartrpchanneltrpy1isinhibitedbyca2calmodulinbinding |