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Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4
Molecular determinants of the binding of various endogenous modulators to transient receptor potential (TRP) channels are crucial for the understanding of necessary cellular pathways, as well as new paths for rational drug designs. The aim of this study was to characterise interactions between the T...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352223/ https://www.ncbi.nlm.nih.gov/pubmed/32560560 http://dx.doi.org/10.3390/ijms21124323 |
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author | Bousova, Kristyna Barvik, Ivan Herman, Petr Hofbauerová, Kateřina Monincova, Lenka Majer, Pavel Zouharova, Monika Vetyskova, Veronika Postulkova, Klara Vondrasek, Jiri |
author_facet | Bousova, Kristyna Barvik, Ivan Herman, Petr Hofbauerová, Kateřina Monincova, Lenka Majer, Pavel Zouharova, Monika Vetyskova, Veronika Postulkova, Klara Vondrasek, Jiri |
author_sort | Bousova, Kristyna |
collection | PubMed |
description | Molecular determinants of the binding of various endogenous modulators to transient receptor potential (TRP) channels are crucial for the understanding of necessary cellular pathways, as well as new paths for rational drug designs. The aim of this study was to characterise interactions between the TRP cation channel subfamily melastatin member 4 (TRPM4) and endogenous intracellular modulators—calcium-binding proteins (calmodulin (CaM) and S100A1) and phosphatidylinositol 4, 5-bisphosphate (PIP(2)). We have found binding epitopes at the N- and C-termini of TRPM4 shared by CaM, S100A1 and PIP(2). The binding affinities of short peptides representing the binding epitopes of N- and C-termini were measured by means of fluorescence anisotropy (FA). The importance of representative basic amino acids and their combinations from both peptides for the binding of endogenous TRPM4 modulators was proved using point alanine-scanning mutagenesis. In silico protein–protein docking of both peptides to CaM and S100A1 and extensive molecular dynamics (MD) simulations enabled the description of key stabilising interactions at the atomic level. Recently solved cryo-Electron Microscopy (EM) structures made it possible to put our findings into the context of the entire TRPM4 channel and to deduce how the binding of these endogenous modulators could allosterically affect the gating of TRPM4. Moreover, both identified binding epitopes seem to be ideally positioned to mediate the involvement of TRPM4 in higher-order hetero-multimeric complexes with important physiological functions. |
format | Online Article Text |
id | pubmed-7352223 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-73522232020-07-21 Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4 Bousova, Kristyna Barvik, Ivan Herman, Petr Hofbauerová, Kateřina Monincova, Lenka Majer, Pavel Zouharova, Monika Vetyskova, Veronika Postulkova, Klara Vondrasek, Jiri Int J Mol Sci Article Molecular determinants of the binding of various endogenous modulators to transient receptor potential (TRP) channels are crucial for the understanding of necessary cellular pathways, as well as new paths for rational drug designs. The aim of this study was to characterise interactions between the TRP cation channel subfamily melastatin member 4 (TRPM4) and endogenous intracellular modulators—calcium-binding proteins (calmodulin (CaM) and S100A1) and phosphatidylinositol 4, 5-bisphosphate (PIP(2)). We have found binding epitopes at the N- and C-termini of TRPM4 shared by CaM, S100A1 and PIP(2). The binding affinities of short peptides representing the binding epitopes of N- and C-termini were measured by means of fluorescence anisotropy (FA). The importance of representative basic amino acids and their combinations from both peptides for the binding of endogenous TRPM4 modulators was proved using point alanine-scanning mutagenesis. In silico protein–protein docking of both peptides to CaM and S100A1 and extensive molecular dynamics (MD) simulations enabled the description of key stabilising interactions at the atomic level. Recently solved cryo-Electron Microscopy (EM) structures made it possible to put our findings into the context of the entire TRPM4 channel and to deduce how the binding of these endogenous modulators could allosterically affect the gating of TRPM4. Moreover, both identified binding epitopes seem to be ideally positioned to mediate the involvement of TRPM4 in higher-order hetero-multimeric complexes with important physiological functions. MDPI 2020-06-17 /pmc/articles/PMC7352223/ /pubmed/32560560 http://dx.doi.org/10.3390/ijms21124323 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Bousova, Kristyna Barvik, Ivan Herman, Petr Hofbauerová, Kateřina Monincova, Lenka Majer, Pavel Zouharova, Monika Vetyskova, Veronika Postulkova, Klara Vondrasek, Jiri Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4 |
title | Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4 |
title_full | Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4 |
title_fullStr | Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4 |
title_full_unstemmed | Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4 |
title_short | Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4 |
title_sort | mapping of cam, s100a1 and pip2-binding epitopes in the intracellular n- and c-termini of trpm4 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352223/ https://www.ncbi.nlm.nih.gov/pubmed/32560560 http://dx.doi.org/10.3390/ijms21124323 |
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