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Title: Arrhythmia mutations in calmodulin cause conformational changes that affect interactions with the cardiac voltage-gated calcium channel

Abstract

Calmodulin (CaM) represents one of the most conserved proteins among eukaryotes and is known to bind and modulate more than a 100 targets. Recently, several disease-associated mutations have been identified in theCALMgenes that are causative of severe cardiac arrhythmia syndromes. Although several mutations have been shown to affect the function of various cardiac ion channels, direct structural insights into any CaM disease mutation have been lacking. Here we report a crystallographic and NMR investigation of several disease mutant CaMs, linked to long-QT syndrome, in complex with the IQ domain of the cardiac voltage-gated calcium channel (Ca V1.2). Surprisingly, two mutants (D95V, N97I) cause a major distortion of the C-terminal lobe, resulting in a pathological conformation not reported before. Furthermore, these structural changes result in altered interactions with the Ca V1.2 IQ domain. Another mutation (N97S) reduces the affinity for Ca 2+by introducing strain in EF hand 3. A fourth mutant (F141L) shows structural changes in the Ca 2+-free state that increase the affinity for the IQ domain. These results thus show that different mechanisms underlie the ability of CaM disease mutations to affect Ca 2+-dependent inactivation of the voltage-gated calcium channel.

Authors:
 [1];  [2];  [1];  [2];  [2]; ORCiD logo [2]; ORCiD logo [2];  [1]
  1. Univ. of British Columbia, Vancouver, BC (Canada)
  2. Aalborg Univ. (Denmark)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
Danish Ministry of Higher Education and Science; Natural Sciences and Engineering Research Council of Canada (NSERC); National Research Council Canada (NRC); Canadian Institutes of Health Research (CIHR); Province of Saskatchewan; Western Economic Diversification Canada; University of Saskatchewan; Obel Family Foundation; Novo Nordisk Foundation; Lundbeck Foundation; Danish Council for Independent Research (DFF); SparNord; Carlsberg Foundations
OSTI Identifier:
1483880
Grant/Contract Number:  
AU-2010-612-181; PJT-148632; NNF15OC0012345; NNF16OC0023344; R151-2013-14432; DFF-4181-00447; DFF-1323-00344; NNF15OC0016186
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 115; Journal Issue: 45; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; calcium signaling; X-ray crystallography; NMR; calcium channels; inactivation

Citation Formats

Wang, Kaiqian, Holt, Christian, Lu, Jocelyn, Brohus, Malene, Larsen, Kamilla Taunsig, Overgaard, Michael Toft, Wimmer, Reinhard, and Van Petegem, Filip. Arrhythmia mutations in calmodulin cause conformational changes that affect interactions with the cardiac voltage-gated calcium channel. United States: N. p., 2018. Web. doi:10.1073/pnas.1808733115.
Wang, Kaiqian, Holt, Christian, Lu, Jocelyn, Brohus, Malene, Larsen, Kamilla Taunsig, Overgaard, Michael Toft, Wimmer, Reinhard, & Van Petegem, Filip. Arrhythmia mutations in calmodulin cause conformational changes that affect interactions with the cardiac voltage-gated calcium channel. United States. doi:10.1073/pnas.1808733115.
Wang, Kaiqian, Holt, Christian, Lu, Jocelyn, Brohus, Malene, Larsen, Kamilla Taunsig, Overgaard, Michael Toft, Wimmer, Reinhard, and Van Petegem, Filip. Mon . "Arrhythmia mutations in calmodulin cause conformational changes that affect interactions with the cardiac voltage-gated calcium channel". United States. doi:10.1073/pnas.1808733115. https://www.osti.gov/servlets/purl/1483880.
@article{osti_1483880,
title = {Arrhythmia mutations in calmodulin cause conformational changes that affect interactions with the cardiac voltage-gated calcium channel},
author = {Wang, Kaiqian and Holt, Christian and Lu, Jocelyn and Brohus, Malene and Larsen, Kamilla Taunsig and Overgaard, Michael Toft and Wimmer, Reinhard and Van Petegem, Filip},
abstractNote = {Calmodulin (CaM) represents one of the most conserved proteins among eukaryotes and is known to bind and modulate more than a 100 targets. Recently, several disease-associated mutations have been identified in theCALMgenes that are causative of severe cardiac arrhythmia syndromes. Although several mutations have been shown to affect the function of various cardiac ion channels, direct structural insights into any CaM disease mutation have been lacking. Here we report a crystallographic and NMR investigation of several disease mutant CaMs, linked to long-QT syndrome, in complex with the IQ domain of the cardiac voltage-gated calcium channel (CaV1.2). Surprisingly, two mutants (D95V, N97I) cause a major distortion of the C-terminal lobe, resulting in a pathological conformation not reported before. Furthermore, these structural changes result in altered interactions with the CaV1.2 IQ domain. Another mutation (N97S) reduces the affinity for Ca2+by introducing strain in EF hand 3. A fourth mutant (F141L) shows structural changes in the Ca2+-free state that increase the affinity for the IQ domain. These results thus show that different mechanisms underlie the ability of CaM disease mutations to affect Ca2+-dependent inactivation of the voltage-gated calcium channel.},
doi = {10.1073/pnas.1808733115},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
issn = {0027-8424},
number = 45,
volume = 115,
place = {United States},
year = {2018},
month = {10}
}

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