Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket

<p dir="ltr">Calmodulin (CaM) is a small, multifunctional calcium (Ca<sup>2+</sup>)-binding sensor that binds and regulates the open probability of cardiac ryanodine receptor 2 (RyR2) at both low and high cytosolic Ca<sup>2+</sup> concentrations. Recent isothe...

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التفاصيل البيبلوغرافية
المؤلف الرئيسي: Angelos Thanassoulas (2822861) (author)
مؤلفون آخرون: Maria Theodoridou (33087) (author), Laila Barrak (17714490) (author), Emna Riguene (17714493) (author), Tamader Alyaarabi (17714496) (author), Mohamed A. Elrayess (7956179) (author), F. Anthony Lai (17714499) (author), Michail Nomikos (17563188) (author)
منشور في: 2023
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_version_ 1864513531028701184
author Angelos Thanassoulas (2822861)
author2 Maria Theodoridou (33087)
Laila Barrak (17714490)
Emna Riguene (17714493)
Tamader Alyaarabi (17714496)
Mohamed A. Elrayess (7956179)
F. Anthony Lai (17714499)
Michail Nomikos (17563188)
author2_role author
author
author
author
author
author
author
author_facet Angelos Thanassoulas (2822861)
Maria Theodoridou (33087)
Laila Barrak (17714490)
Emna Riguene (17714493)
Tamader Alyaarabi (17714496)
Mohamed A. Elrayess (7956179)
F. Anthony Lai (17714499)
Michail Nomikos (17563188)
author_role author
dc.creator.none.fl_str_mv Angelos Thanassoulas (2822861)
Maria Theodoridou (33087)
Laila Barrak (17714490)
Emna Riguene (17714493)
Tamader Alyaarabi (17714496)
Mohamed A. Elrayess (7956179)
F. Anthony Lai (17714499)
Michail Nomikos (17563188)
dc.date.none.fl_str_mv 2023-10-26T03:00:00Z
dc.identifier.none.fl_str_mv 10.3390/ijms242115630
dc.relation.none.fl_str_mv https://figshare.com/articles/journal_contribution/Arrhythmia-Associated_Calmodulin_E105A_Mutation_Alters_the_Binding_Affinity_of_CaM_to_a_Ryanodine_Receptor_2_CaM-Binding_Pocket/24921522
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biomedical and clinical sciences
Cardiovascular medicine and haematology
Medical physiology
Chemical sciences
Medicinal and biomolecular chemistry
calmodulin
ryanodine receptor
RyR2
arrhythmias
cardiac disease
dc.title.none.fl_str_mv Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
dc.type.none.fl_str_mv Text
Journal contribution
info:eu-repo/semantics/publishedVersion
text
contribution to journal
description <p dir="ltr">Calmodulin (CaM) is a small, multifunctional calcium (Ca<sup>2+</sup>)-binding sensor that binds and regulates the open probability of cardiac ryanodine receptor 2 (RyR2) at both low and high cytosolic Ca<sup>2+</sup> concentrations. Recent isothermal titration calorimetry (ITC) studies of a number of peptides that correspond to different regions of human RyR2 showed that two regions of human RyR2 (3584-3602aa and 4255-4271aa) bind with high affinity to CaM, suggesting that these two regions might contribute to a putative RyR2 intra-subunit CaM-binding pocket. Moreover, a previously characterized de novo long QT syndrome (LQTS)-associated missense CaM mutation (E105A) which was identified in a 6-year-old boy, who experienced an aborted first episode of cardiac arrest revealed that this mutation dysregulates normal cardiac function in zebrafish by a complex mechanism that involves alterations in both CaM-Ca<sup>2+</sup> and CaM-RyR2 interactions. Herein, to gain further insight into how the CaM E105A mutation leads to severe cardiac arrhythmia, we generated large quantities of recombinant CaMWT and CaME105A proteins. We then performed ITC experiments to investigate and compare the interactions of CaMWT and CaME105A mutant protein with two synthetic peptides that correspond to the two aforementioned human RyR2 regions, which we have proposed to contribute to the RyR2 CaM-binding pocket. Our data reveal that the E105A mutation has a significant negative effect on the interaction of CaM with both RyR2 regions in the presence and absence of Ca<sup>2+</sup>, highlighting the potential contribution of these two human RyR2 regions to an RyR2 CaM-binding pocket, which may be essential for physiological CaM/RyR2 association and thus channel regulation.</p><h2>Other Information</h2><p dir="ltr">Published in: International Journal of Molecular Sciences<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/ijms242115630" target="_blank">https://dx.doi.org/10.3390/ijms242115630</a></p>
eu_rights_str_mv openAccess
id Manara2_9a400e3f69e349a5bdb810547bb027fa
identifier_str_mv 10.3390/ijms242115630
network_acronym_str Manara2
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publishDate 2023
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spelling Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding PocketAngelos Thanassoulas (2822861)Maria Theodoridou (33087)Laila Barrak (17714490)Emna Riguene (17714493)Tamader Alyaarabi (17714496)Mohamed A. Elrayess (7956179)F. Anthony Lai (17714499)Michail Nomikos (17563188)Biomedical and clinical sciencesCardiovascular medicine and haematologyMedical physiologyChemical sciencesMedicinal and biomolecular chemistrycalmodulinryanodine receptorRyR2arrhythmiascardiac disease<p dir="ltr">Calmodulin (CaM) is a small, multifunctional calcium (Ca<sup>2+</sup>)-binding sensor that binds and regulates the open probability of cardiac ryanodine receptor 2 (RyR2) at both low and high cytosolic Ca<sup>2+</sup> concentrations. Recent isothermal titration calorimetry (ITC) studies of a number of peptides that correspond to different regions of human RyR2 showed that two regions of human RyR2 (3584-3602aa and 4255-4271aa) bind with high affinity to CaM, suggesting that these two regions might contribute to a putative RyR2 intra-subunit CaM-binding pocket. Moreover, a previously characterized de novo long QT syndrome (LQTS)-associated missense CaM mutation (E105A) which was identified in a 6-year-old boy, who experienced an aborted first episode of cardiac arrest revealed that this mutation dysregulates normal cardiac function in zebrafish by a complex mechanism that involves alterations in both CaM-Ca<sup>2+</sup> and CaM-RyR2 interactions. Herein, to gain further insight into how the CaM E105A mutation leads to severe cardiac arrhythmia, we generated large quantities of recombinant CaMWT and CaME105A proteins. We then performed ITC experiments to investigate and compare the interactions of CaMWT and CaME105A mutant protein with two synthetic peptides that correspond to the two aforementioned human RyR2 regions, which we have proposed to contribute to the RyR2 CaM-binding pocket. Our data reveal that the E105A mutation has a significant negative effect on the interaction of CaM with both RyR2 regions in the presence and absence of Ca<sup>2+</sup>, highlighting the potential contribution of these two human RyR2 regions to an RyR2 CaM-binding pocket, which may be essential for physiological CaM/RyR2 association and thus channel regulation.</p><h2>Other Information</h2><p dir="ltr">Published in: International Journal of Molecular Sciences<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/ijms242115630" target="_blank">https://dx.doi.org/10.3390/ijms242115630</a></p>2023-10-26T03:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.3390/ijms242115630https://figshare.com/articles/journal_contribution/Arrhythmia-Associated_Calmodulin_E105A_Mutation_Alters_the_Binding_Affinity_of_CaM_to_a_Ryanodine_Receptor_2_CaM-Binding_Pocket/24921522CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/249215222023-10-26T03:00:00Z
spellingShingle Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
Angelos Thanassoulas (2822861)
Biomedical and clinical sciences
Cardiovascular medicine and haematology
Medical physiology
Chemical sciences
Medicinal and biomolecular chemistry
calmodulin
ryanodine receptor
RyR2
arrhythmias
cardiac disease
status_str publishedVersion
title Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
title_full Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
title_fullStr Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
title_full_unstemmed Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
title_short Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
title_sort Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket
topic Biomedical and clinical sciences
Cardiovascular medicine and haematology
Medical physiology
Chemical sciences
Medicinal and biomolecular chemistry
calmodulin
ryanodine receptor
RyR2
arrhythmias
cardiac disease