The ryanodine receptor (RyR) is a Ca2+ release channel that is present in the sarcoplasmic reticulum and endoplasmic reticulum (ER) and that plays a central role in excitation-contraction coupling in skeletal and cardiac muscle. Hyperactivation of RyR by genetic mutations or posttranslational modification can cause various skeletal muscle and arrhythmogenic heart diseases. Inhibitors of RyR are therefore expected to be potential drugs for treatment of such diseases. This article describes assays to evaluate RyR channel activity, including an ER Ca2+ measurement assay that is compatible with high-throughput screening and a [3 H]-ryanodine binding assay that provides a quantitative measure of RyR channel activity as a second screen for compound hits.
The ryanodine receptor (RyR) is a Ca2+ release channel that is present in the sarcoplasmic reticulum and endoplasmic reticulum (ER) and that plays a central role in excitation-contraction coupling in skeletal and cardiac muscle. Hyperactivation of RyR by genetic mutations or posttranslational modification can cause various skeletal muscle and arrhythmogenic heart diseases. Inhibitors of RyR are therefore expected to be potential drugs for treatment of such diseases. This article describes assays to evaluate RyR channel activity, including an ER Ca2+ measurement assay that is compatible with high-throughput screening and a [3 H]-ryanodine binding assay that provides a quantitative measure of RyR channel activity as a second screen for compound hits.