Unique phosphorylation site on the cardiac ryanodine receptor regulates calcium channel activity.
Ryanodine receptors have recently been shown to be the Ca2+ release channels of sarcoplasmic reticulum in both cardiac muscle and skeletal muscle. Several regulatory sites are postulated to exist on these receptors, but to date, none have been definitively identified. In the work described here, we localize one of these sites by showing that the cardiac isoform of the ryanodine receptor is a preferred substrate for multifunctional Ca2+/calmodulin-dependent protein kinase (CaM kinase). Phosphorylation by CaM kinase occurs at a single site encompassing serine 2809. Antibodies generated to this site react only with the cardiac isoform of the ryanodine receptor, and immunoprecipitate only cardiac [3H]ryanodine-binding sites. When cardiac junctional sarcoplasmic reticulum vesicles or partially purified ryanodine receptors are fused with planar bilayers, phosphorylation at this site activates the Ca2+ channel. In tissues expressing the cardiac isoform of the ryanodine receptor, such as heart and brain, phosphorylation of the Ca2+ release channel by CaM kinase may provide a unique mechanism for regulating intracellular Ca2+ release.
Witcher
D R
DR
Krannert Institute of Cardiology, Indiana University School of Medicine, Indianapolis 46202.
Kovacs
R J
RJ
Schulman
H
H
Cefali
D C
DC
Jones
L R
LR
D000595:Amino Acid Sequence; D000818:Animals; D015220:Calcium Channels / Q000502:physiology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D004285:Dogs; D006321:Heart / Q000502:physiology*; D015151:Immunoblotting; D007700:Kinetics; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D009132:Muscles / Q000502:physiology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D012433:Ryanodine / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel; D012519:Sarcoplasmic Reticulum / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
D015220:Calcium Channels; D011950:Receptors, Cholinergic; D019837:Ryanodine Receptor Calcium Release Channel; D012433:Ryanodine; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases