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Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium

BACKGROUND: Calmodulin (CaM) is an evolutionarily conserved eukaryotic multifunctional protein that functions as the major sensor of intracellular calcium signaling. Its calcium-modulated function regulates the activity of numerous effector proteins involved in a variety of physiological processes i...

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Autores principales: Léger, Corentin, Pitard, Irène, Sadi, Mirko, Carvalho, Nicolas, Brier, Sébastien, Mechaly, Ariel, Raoux-Barbot, Dorothée, Davi, Maryline, Hoos, Sylviane, Weber, Patrick, Vachette, Patrice, Durand, Dominique, Haouz, Ahmed, Guijarro, J. Iñaki, Ladant, Daniel, Chenal, Alexandre
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9361521/
https://www.ncbi.nlm.nih.gov/pubmed/35945584
http://dx.doi.org/10.1186/s12915-022-01381-5
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author Léger, Corentin
Pitard, Irène
Sadi, Mirko
Carvalho, Nicolas
Brier, Sébastien
Mechaly, Ariel
Raoux-Barbot, Dorothée
Davi, Maryline
Hoos, Sylviane
Weber, Patrick
Vachette, Patrice
Durand, Dominique
Haouz, Ahmed
Guijarro, J. Iñaki
Ladant, Daniel
Chenal, Alexandre
author_facet Léger, Corentin
Pitard, Irène
Sadi, Mirko
Carvalho, Nicolas
Brier, Sébastien
Mechaly, Ariel
Raoux-Barbot, Dorothée
Davi, Maryline
Hoos, Sylviane
Weber, Patrick
Vachette, Patrice
Durand, Dominique
Haouz, Ahmed
Guijarro, J. Iñaki
Ladant, Daniel
Chenal, Alexandre
author_sort Léger, Corentin
collection PubMed
description BACKGROUND: Calmodulin (CaM) is an evolutionarily conserved eukaryotic multifunctional protein that functions as the major sensor of intracellular calcium signaling. Its calcium-modulated function regulates the activity of numerous effector proteins involved in a variety of physiological processes in diverse organs, from proliferation and apoptosis, to memory and immune responses. Due to the pleiotropic roles of CaM in normal and pathological cell functions, CaM antagonists are needed for fundamental studies as well as for potential therapeutic applications. Calmidazolium (CDZ) is a potent small molecule antagonist of CaM and one the most widely used inhibitors of CaM in cell biology. Yet, CDZ, as all other CaM antagonists described thus far, also affects additional cellular targets and its lack of selectivity hinders its application for dissecting calcium/CaM signaling. A better understanding of CaM:CDZ interaction is key to design analogs with improved selectivity. Here, we report a molecular characterization of CaM:CDZ complexes using an integrative structural biology approach combining SEC-SAXS, X-ray crystallography, HDX-MS, and NMR. RESULTS: We provide evidence that binding of a single molecule of CDZ induces an open-to-closed conformational reorientation of the two domains of CaM and results in a strong stabilization of its structural elements associated with a reduction of protein dynamics over a large time range. These CDZ-triggered CaM changes mimic those induced by CaM-binding peptides derived from physiological protein targets, despite their distinct chemical natures. CaM residues in close contact with CDZ and involved in the stabilization of the CaM:CDZ complex have been identified. CONCLUSION: Our results provide molecular insights into CDZ-induced dynamics and structural changes of CaM leading to its inhibition and open the way to the rational design of more selective CaM antagonists. GRAPHICAL ABSTRACT: Calmidazolium is a potent and widely used inhibitor of calmodulin, a major mediator of calcium-signaling in eukaryotic cells. Structural characterization of calmidazolium-binding to calmodulin reveals that it triggers open-to-closed conformational changes similar to those induced by calmodulin-binding peptides derived from enzyme targets. These results provide molecular insights into CDZ-induced dynamics and structural changes of CaM leading to its inhibition and open the way to the rational design of more selective CaM antagonists. [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12915-022-01381-5.
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spelling pubmed-93615212022-08-10 Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium Léger, Corentin Pitard, Irène Sadi, Mirko Carvalho, Nicolas Brier, Sébastien Mechaly, Ariel Raoux-Barbot, Dorothée Davi, Maryline Hoos, Sylviane Weber, Patrick Vachette, Patrice Durand, Dominique Haouz, Ahmed Guijarro, J. Iñaki Ladant, Daniel Chenal, Alexandre BMC Biol Research Article BACKGROUND: Calmodulin (CaM) is an evolutionarily conserved eukaryotic multifunctional protein that functions as the major sensor of intracellular calcium signaling. Its calcium-modulated function regulates the activity of numerous effector proteins involved in a variety of physiological processes in diverse organs, from proliferation and apoptosis, to memory and immune responses. Due to the pleiotropic roles of CaM in normal and pathological cell functions, CaM antagonists are needed for fundamental studies as well as for potential therapeutic applications. Calmidazolium (CDZ) is a potent small molecule antagonist of CaM and one the most widely used inhibitors of CaM in cell biology. Yet, CDZ, as all other CaM antagonists described thus far, also affects additional cellular targets and its lack of selectivity hinders its application for dissecting calcium/CaM signaling. A better understanding of CaM:CDZ interaction is key to design analogs with improved selectivity. Here, we report a molecular characterization of CaM:CDZ complexes using an integrative structural biology approach combining SEC-SAXS, X-ray crystallography, HDX-MS, and NMR. RESULTS: We provide evidence that binding of a single molecule of CDZ induces an open-to-closed conformational reorientation of the two domains of CaM and results in a strong stabilization of its structural elements associated with a reduction of protein dynamics over a large time range. These CDZ-triggered CaM changes mimic those induced by CaM-binding peptides derived from physiological protein targets, despite their distinct chemical natures. CaM residues in close contact with CDZ and involved in the stabilization of the CaM:CDZ complex have been identified. CONCLUSION: Our results provide molecular insights into CDZ-induced dynamics and structural changes of CaM leading to its inhibition and open the way to the rational design of more selective CaM antagonists. GRAPHICAL ABSTRACT: Calmidazolium is a potent and widely used inhibitor of calmodulin, a major mediator of calcium-signaling in eukaryotic cells. Structural characterization of calmidazolium-binding to calmodulin reveals that it triggers open-to-closed conformational changes similar to those induced by calmodulin-binding peptides derived from enzyme targets. These results provide molecular insights into CDZ-induced dynamics and structural changes of CaM leading to its inhibition and open the way to the rational design of more selective CaM antagonists. [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12915-022-01381-5. BioMed Central 2022-08-09 /pmc/articles/PMC9361521/ /pubmed/35945584 http://dx.doi.org/10.1186/s12915-022-01381-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Léger, Corentin
Pitard, Irène
Sadi, Mirko
Carvalho, Nicolas
Brier, Sébastien
Mechaly, Ariel
Raoux-Barbot, Dorothée
Davi, Maryline
Hoos, Sylviane
Weber, Patrick
Vachette, Patrice
Durand, Dominique
Haouz, Ahmed
Guijarro, J. Iñaki
Ladant, Daniel
Chenal, Alexandre
Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium
title Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium
title_full Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium
title_fullStr Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium
title_full_unstemmed Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium
title_short Dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium
title_sort dynamics and structural changes of calmodulin upon interaction with the antagonist calmidazolium
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9361521/
https://www.ncbi.nlm.nih.gov/pubmed/35945584
http://dx.doi.org/10.1186/s12915-022-01381-5
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