Mutation-linked, excessively tight interaction between the calmodulin binding domain and the C-terminal domain of the cardiac ryanodine receptor as a novel cause of catecholaminergic polymorphic ventricular tachycardia.

Abstract

BACKGROUND:

Ryanodine receptor (RyR2) is known to be a causal gene of catecholaminergic polymorphic ventricular tachycardia (CPVT), an important inherited disease. Some of the human CPVT-associated mutations have been found in a domain (4026-4172) that has EF hand motifs, the so-called calmodulin (CaM)-like domain (CaMLD).

OBJECTIVE:

The purpose of this study was to investigate the underlying mechanism by which CPVT is induced by a mutation at CaMLD.

METHODS:

A new N4103K/+ knock-in (KI) mice model was generated.

RESULTS:

Sustained ventricular tachycardia was frequently observed after infusion of caffeine plus epinephrine in KI mice. Endogenous CaM bound to RyR2 decreased even at baseline in isolated KI cardiomyocytes. Ca2+ spark frequency (CaSpF) was much higher in KI cells than in wild-type cells. Addition of GSH-CaM (higher affinity CaM to RyR2) significantly decreased CaSpF. In response to isoproterenol, spontaneous Ca2+ transient (SCaT) was frequently observed in intact KI cells. Incorporation of GSH-CaM into intact KI cells using a protein delivery kit decreased SCaT significantly. An assay using a quartz crystal microbalance technique revealed that mutated CaMLD peptide showed higher binding affinity to CaM binding domain (CaMBD) peptide.

CONCLUSION:

In the N4103K mutant, CaM binding affinity to RyR2 was significantly reduced regardless of beta-adrenergic stimulation. We found that this was caused by an abnormally tight interaction between CaMBD and mutated CaM-like domain (N4103K-CaMBD). Thus, CaMBD-CaMLD interaction may be a novel therapeutic target for treatment of lethal arrhythmia.

• Authors: Shigehiko Nishimura, Takeshi Yamamoto, Yoshihide Nakamura, Michiaki Kohno, Yoriomi Hamada, Yoko Sufu, Go Fukui, Takuma Nanno, Hironori Ishiguchi, Takayoshi Kato, Xiaojuan Xu, Makoto Ono, Tetsuro Oda, Shinichi Okuda, Shigeki Kobayashi, Masafumi Yano
• Journal: Heart rhythm (Heart Rhythm) Vol. 15 Issue 6 Pg. 905-914 (06 2018) ISSN: 1556-3871 [Electronic] United States
• PMID: 29427818 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2018 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.

Chemical References

• Ryanodine Receptor Calcium Release Channel
• DNA
• Calcium

Topics

• Animals
• Calcium (metabolism)
• Calcium Signaling
• Cells, Cultured
• DNA (genetics)
• DNA Mutational Analysis
• Disease Models, Animal
• Mice
• Mutation
• Myocytes, Cardiac (metabolism, pathology)
• Ryanodine Receptor Calcium Release Channel (genetics, metabolism)
• Tachycardia, Ventricular (genetics, metabolism, pathology)