Defective calmodulin binding to the cardiac ryanodine receptor plays a key role in CPVT-associated channel dysfunction.

Abstract

Calmodulin (CaM), one of the accessory proteins of the cardiac ryanodine receptor (RyR2), is known to play a significant role in the channel regulation of the RyR2. However, the possible involvement of calmodulin in the pathogenic process of catecholaminergic polymorphic ventricular tachycardia (CPVT) has not been investigated. In this study, we investigated the state of RyR2-bound CaM and channel dysfunctions using a knock-in (KI) mouse model with CPVT-linked RyR2 mutation (R2474S). Without added effectors, the affinity of CaM binding to the RyR2 was indistinguishable between KI and WT hearts. In response to cAMP (1 micromol/L), the RyR2 phosphorylation at Ser2808 increased in both WT and KI hearts to the same extent. However, cAMP caused a significant decrease of the CaM-binding affinity in KI hearts, but the affinity was unchanged in WT. Dantrolene restored a normal level of CaM-binding affinity in the cAMP-treated KI hearts, suggesting that defective inter-domain interaction between the N-terminal domain and the central domain of the RyR2 (the target of therapeutic effect of dantrolene) is involved in the cAMP-induced reduction of the CaM-binding affinity. In saponin-permeabilized cardiomyocytes, the addition of cAMP increased the frequency of spontaneous Ca(2+) sparks to a significantly larger extent in KI cardiomyocytes than in WT cardiomyocytes, whereas the addition of a high concentration of CaM attenuated the aberrant increase of Ca(2+) sparks. In conclusion, CPVT mutation causes defective inter-domain interaction, significant reduction in the ability of CaM binding to the RyR2, spontaneous Ca(2+) leak, and then lethal arrhythmia.

• Authors: Xiaojuan Xu, Masafumi Yano, Hitoshi Uchinoumi, Akihiro Hino, Takeshi Suetomi, Makoto Ono, Hiroki Tateishi, Tetsuro Oda, Shinichi Okuda, Masahiro Doi, Shigeki Kobayashi, Takeshi Yamamoto, Yasuhiro Ikeda, Noriaki Ikemoto, Masunori Matsuzaki
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 394 Issue 3 Pg. 660-6 (Apr 09 2010) ISSN: 1090-2104 [Electronic] United States
• PMID: 20226167 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 2010 Elsevier Inc. All rights reserved.

Chemical References

• Calmodulin
• Ryanodine Receptor Calcium Release Channel
• Cyclic AMP-Dependent Protein Kinases
• Calcium

Topics

• Animals
• Calcium (metabolism)
• Calmodulin (metabolism)
• Cyclic AMP-Dependent Protein Kinases (metabolism)
• Gene Knock-In Techniques
• Mice
• Mice, Mutant Strains
• Mutation
• Myocytes, Cardiac (metabolism)
• Ryanodine Receptor Calcium Release Channel (genetics, metabolism)
• Tachycardia, Ventricular (genetics, metabolism)