%0 Journal Article %1 Zhao_2001_13810 %A Zhao, F. %A Li, P. %A Chen, S. R. %A Louis, C. F. %A Fruen, B. R. %D 2001 %J J. Biol. Chem. %K 11278295 50, Acid, Adenine, Adenosine Alanine, Animals, Arginine, Binding Binding, Caffeine, Calcium Calcium, Cell Central Central, Channel Channel, Chloride, Cloning, Co, Complementary, Concentration Conformation, Cryoelectron Cysteine, DNA, Dantrolene, Dominant, Dose-Response Drug, Electrophysiology, Fusion Gating, Genes, Gl, Glutamic Glutathione Gov't, Heart, Humans, Inhibitory Ion Isoforms, Kinetics, Line, Magnesium Magnesium, Mice, Microscopy, Molecular, Muscle Muscle, Mutation, Myocardium, Nervous Non-U.S. P.H.S., Point Protein Proteins, Receptor Recombinant Relationship, Relaxants, Release Research Reticulum, Ryanodine Ryanodine, Sarcoplasmic Signal Sites, Skeletal, Stimulants, Strontium, Support, Swine, System Transduction, Transfection, Transferase, Triphosphate, U.S. ntraction, utamic %N 17 %P 13810--13816 %R 10.1074/jbc.M006104200 %T Dantrolene inhibition of ryanodine receptor Ca$^2+$ release channels. Molecular mechanism and isoform selectivity. %U http://dx.doi.org/10.1074/jbc.M006104200 %V 276 %X As an inhibitor of Ca$^2+$ release through ryanodine receptor (RYR) channels, the skeletal muscle relaxant dantrolene has proven to be both a valuable experimental probe of intracellular Ca$^2+$ signaling and a lifesaving treatment for the pharmacogenetic disorder malignant hyperthermia. However, the molecular basis and specificity of the actions of dantrolene on RYR channels have remained in question. Here we utilize (3)Hryanodine binding to further investigate the actions of dantrolene on the three mammalian RYR isoforms. The inhibition of the pig skeletal muscle RYR1 by dantrolene (10 microm) was associated with a 3-fold increase in the K(d) of (3)Hryanodine binding to sarcoplasmic reticulum (SR) vesicles such that dantrolene effectively reversed the 3-fold decrease in the K(d) for (3)Hryanodine binding resulting from the malignant hyperthermia RYR1 Arg(615) --> Cys mutation. Dantrolene inhibition of the RYR1 was dependent on the presence of the adenine nucleotide and calmodulin and reflected a selective decrease in the apparent affinity of RYR1 activation sites for Ca$^2+$ relative to Mg$^2+$. In contrast to the RYR1 isoform, the cardiac RYR2 isoform was unaffected by dantrolene, both in native cardiac SR vesicles and when heterologously expressed in HEK-293 cells. By comparison, the RYR3 isoform expressed in HEK-293 cells was significantly inhibited by dantrolene, and the extent of RYR3 inhibition was similar to that displayed by the RYR1 in native SR vesicles. Our results thus indicate that both the RYR1 and the RYR3, but not the RYR2, may be targets for dantrolene inhibition in vivo.

@article{Zhao_2001_13810, abstract = {As an inhibitor of {C}a$^{2+}$ release through ryanodine receptor (RYR) channels, the skeletal muscle relaxant dantrolene has proven to be both a valuable experimental probe of intracellular {C}a$^{2+}$ signaling and a lifesaving treatment for the pharmacogenetic disorder malignant hyperthermia. However, the molecular basis and specificity of the actions of dantrolene on RYR channels have remained in question. Here we utilize [(3)H]ryanodine binding to further investigate the actions of dantrolene on the three mammalian RYR isoforms. The inhibition of the pig skeletal muscle RYR1 by dantrolene (10 microm) was associated with a 3-fold increase in the K(d) of [(3)H]ryanodine binding to sarcoplasmic reticulum (SR) vesicles such that dantrolene effectively reversed the 3-fold decrease in the K(d) for [(3)H]ryanodine binding resulting from the malignant hyperthermia RYR1 Arg(615) --> Cys mutation. Dantrolene inhibition of the RYR1 was dependent on the presence of the adenine nucleotide and calmodulin and reflected a selective decrease in the apparent affinity of RYR1 activation sites for {C}a$^{2+}$ relative to {M}g$^{2+}$. In contrast to the RYR1 isoform, the cardiac RYR2 isoform was unaffected by dantrolene, both in native cardiac SR vesicles and when heterologously expressed in HEK-293 cells. By comparison, the RYR3 isoform expressed in HEK-293 cells was significantly inhibited by dantrolene, and the extent of RYR3 inhibition was similar to that displayed by the RYR1 in native SR vesicles. Our results thus indicate that both the RYR1 and the RYR3, but not the RYR2, may be targets for dantrolene inhibition in vivo.}, added-at = {2009-06-03T11:20:58.000+0200}, author = {Zhao, F. and Li, P. and Chen, S. R. and Louis, C. F. and Fruen, B. R.}, biburl = {https://www.bibsonomy.org/bibtex/2a709303295845de834cceb11950272ae/hake}, description = {The whole bibliography file I use.}, doi = {10.1074/jbc.M006104200}, interhash = {3fd4fcb34797accb005eef4fa23a44ef}, intrahash = {a709303295845de834cceb11950272ae}, journal = {J. Biol. Chem.}, keywords = {11278295 50, Acid, Adenine, Adenosine Alanine, Animals, Arginine, Binding Binding, Caffeine, Calcium Calcium, Cell Central Central, Channel Channel, Chloride, Cloning, Co, Complementary, Concentration Conformation, Cryoelectron Cysteine, DNA, Dantrolene, Dominant, Dose-Response Drug, Electrophysiology, Fusion Gating, Genes, Gl, Glutamic Glutathione Gov't, Heart, Humans, Inhibitory Ion Isoforms, Kinetics, Line, Magnesium Magnesium, Mice, Microscopy, Molecular, Muscle Muscle, Mutation, Myocardium, Nervous Non-U.S. P.H.S., Point Protein Proteins, Receptor Recombinant Relationship, Relaxants, Release Research Reticulum, Ryanodine Ryanodine, Sarcoplasmic Signal Sites, Skeletal, Stimulants, Strontium, Support, Swine, System Transduction, Transfection, Transferase, Triphosphate, U.S. ntraction, utamic}, month = Apr, number = 17, pages = {13810--13816}, pii = {M006104200}, pmid = {11278295}, timestamp = {2009-06-03T11:21:39.000+0200}, title = {Dantrolene inhibition of ryanodine receptor {C}a$^{2+}$ release channels. Molecular mechanism and isoform selectivity.}, url = {http://dx.doi.org/10.1074/jbc.M006104200}, volume = 276, year = 2001 }