Ion channel gating in cardiac ryanodine receptors from the arrhythmic RyR2-P2328S mouse

ABSTRACT

Mutations in the cardiac ryanodine receptor Ca ²⁺ release channel (RyR2) can cause deadly ventricular arrhythmias and atrial fibrillation (AF). The RyR2-P2328S mutation produces catecholaminergic polymorphic ventricular tachycardia (CPVT) and AF in hearts from homozygous RyR2 ^{P2328S/P2328S} (denoted RyR2 ^S/S) mice. We have now examined P2328S RyR2 channels from RyR2 ^S/S hearts. The activity of wild-type (WT) and P2328S RyR2 channels was similar at a cytoplasmic [Ca ²⁺] of 1 mM, but P2328S RyR2 was significantly more active than WT at a cytoplasmic [Ca ²⁺] of 1 µM. This was associated with a >10-fold shift in the half maximal activation concentration (AC ₅₀) for Ca ²⁺ activation, from ∼3.5 µM Ca ²⁺ in WT RyR2 to ∼320 nM in P2328S channels and an unexpected >1000-fold shift in the half maximal inhibitory concentration (IC ₅₀) for inactivation from ∼50 mM in WT channels to ≤7 μM in P2328S channels, which is into systolic [Ca ²⁺] levels. Unexpectedly, the shift in Ca ²⁺ activation was not associated with changes in sub-conductance activity, S2806 or S2814 phosphorylation or the level of FKBP12 (also known as FKBP1A) bound to the channels. The changes in channel activity seen with the P2328S mutation correlate with altered Ca ²⁺ homeostasis in myocytes from RyR2 ^S/S mice and the CPVT and AF phenotypes.

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Abstract

Summary: The RyR2–P2328S mutation, precipitating potentially fatal arrhythmia in humans, can be attributed to leftward shifts in cytoplasmic Ca ²⁺–dependent RyR2 activation and inactivation in the absence of adrenergic stimulation.