European Biophysics Journal

, Volume 37, Issue 4, pp 455–467 | Cite as

A domain peptide of the cardiac ryanodine receptor regulates channel sensitivity to luminal Ca2+ via cytoplasmic Ca2+ sites

  • Derek R. Laver
  • Bonny N. Honen
  • Graham D. Lamb
  • Noriaki Ikemoto
Original Paper

Abstract

The clustering of cardiac RyR mutations, linked to sudden cardiac death (SCD), into several regions in the amino acid sequence underlies the hypothesis that these mutations interfere with stabilising interactions between different domains of the RyR2. SCD mutations cause increased channel sensitivity to cytoplasmic and luminal Ca2+. A synthetic peptide corresponding to part of the central domain (DPc10:2460G–P2495) was designed to destabilise the interaction of the N-terminal and central domains of wild-type RyR2 and mimic the effects of SCD mutations. With Ca2+ as the sole regulating ion, DPc10 caused increased channel activity which could be reversed by removal of the peptide whereas in the presence of ATP DPc10 caused no activation. In support of the domain destablising hypothesis, the corresponding peptide (DPc10-mut) containing the CPVT mutation R2474S did not affect channel activity under any circumstances. DPc10-induced activation was due to a small increase in RyR2 sensitivity to cytoplasmic Ca2+ and a large increase in the magnitude of luminal Ca2+ activation. The increase in the luminal Ca2+ response appeared reliant on the luminal-to-cytoplasmic Ca2+ flux in the channel, indicating that luminal Ca2+ was activating the RyR2 via its cytoplasmic Ca2+ sites. DPc10 had no significant effect on the RyR2 gating associated with luminal Ca2+ sensing sites. The results were fitted by the luminal-triggered Ca2+ feed-through model and the effects of DPc10 were explained entirely by perturbations in cytoplasmic Ca2+-activation mechanism.

Keywords

Sudden cardiac death Calcium release channels Cardiac muscle Calcium stores Excitation–contraction coupling Ryanodine receptor Bilayer 

Abbreviations

SCD

Sudden cardiac death

CPVT

Catecholaminergic polymorphic ventricular tachycardia

ARVD2

Arrhythmogenic right ventricular cardiomyopathy type 2

SR

Sarcoplasmic reticulum

RyR

Ryanodine receptor

RyR2

Cardiac ryanodine receptors

[Ca2+]L

Luminal Ca2+ concentration

[Ca2+]C

Cytoplasmic Ca2+ concentration

TES

N-tris[Hydroxymethyl]methyl-2-aminoethanesulfonic acid

BAPTA

(1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′- tetraacetic acid (4K+)

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Copyright information

© EBSA 2007

Authors and Affiliations

  • Derek R. Laver
    • 1
  • Bonny N. Honen
    • 1
  • Graham D. Lamb
    • 3
  • Noriaki Ikemoto
    • 2
  1. 1.School of Biomedical SciencesUniversity of Newcastle and Hunter Medical Research InstituteCallaghanAustralia
  2. 2.Boston Biomedical Research InstituteWatertownUSA
  3. 3.Department of ZoologyLa Trobe UniversityMelbourneAustralia

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