The cardiac Ca2+ release channel [ryanodine receptor type 2 (RyR2)] is modulated by thiol reactive agents, but the molecular basis of RyR2 modulation by thiol reagents is poorly understood. Cys3635 in the skeletal muscle RyR1 is one of the most hyper-reactive thiols and is important for the redox and calmodulin (CaM) regulation of the RyR1 channel. However, little is known about the role of the corresponding cysteine residue in RyR2 (Cys3602) in the function and regulation of the RyR2 channel. In the present study, we assessed the impact of mutating Cys3602 (C3602A) on store overload-induced Ca2+ release (SOICR) and the regulation of RyR2 by thiol reagents and CaM. We found that the C3602A mutation suppressed SOICR by raising the activation threshold and delayed the termination of Ca2+ release by reducing the termination threshold. As a result, C3602A markedly increased the fractional Ca2+ release. Furthermore, the C3602A mutation diminished the inhibitory effect of N-ethylmaleimide on Ca2+ release, but it had no effect on the stimulatory action of 4,4′-dithiodipyridine (DTDP) on Ca2+ release. In addition, Cys3602 mutations (C3602A or C3602R) did not abolish the effect of CaM on Ca2+-release termination. Therefore, RyR2–Cys3602 is a major site mediating the action of thiol alkylating agent N-ethylmaleimide, but not the action of the oxidant DTDP. Our data also indicate that residue Cys3602 plays an important role in the activation and termination of Ca2+ release, but it is not essential for CaM regulation of RyR2.
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Research Article|
March 20 2015
Role of Cys3602 in the function and regulation of the cardiac ryanodine receptor
Tao Mi;
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Zhichao Xiao;
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Wenting Guo;
Wenting Guo
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Yijun Tang;
Yijun Tang
4
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Florian Hiess;
Florian Hiess
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Jianmin Xiao;
Jianmin Xiao
5
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Yundi Wang;
Yundi Wang
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Joe Z. Zhang;
Joe Z. Zhang
†Department of Physiology and Heart Otago, University of Otago, Dunedin 9045, New Zealand
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Lin Zhang;
Lin Zhang
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Ruiwu Wang;
Ruiwu Wang
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
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Peter P. Jones;
Peter P. Jones
†Department of Physiology and Heart Otago, University of Otago, Dunedin 9045, New Zealand
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S. R. Wayne Chen
*The Libin Cardiovascular Institute of Alberta, Department of Physiology and Pharmacology and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada
7To whom correspondence should be addressed (email swchen@ucalgary.ca).
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Publisher: Portland Press Ltd
Received:
October 09 2014
Revision Received:
January 20 2015
Accepted:
January 21 2015
Accepted Manuscript online:
January 21 2015
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2015 Biochemical Society
2015
Biochem J (2015) 467 (1): 177–190.
Article history
Received:
October 09 2014
Revision Received:
January 20 2015
Accepted:
January 21 2015
Accepted Manuscript online:
January 21 2015
Citation
Tao Mi, Zhichao Xiao, Wenting Guo, Yijun Tang, Florian Hiess, Jianmin Xiao, Yundi Wang, Joe Z. Zhang, Lin Zhang, Ruiwu Wang, Peter P. Jones, S. R. Wayne Chen; Role of Cys3602 in the function and regulation of the cardiac ryanodine receptor. Biochem J 1 April 2015; 467 (1): 177–190. doi: https://doi.org/10.1042/BJ20141263
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