The point mutation Arg615-->Cys in the Ca2+ release channel of skeletal sarcoplasmic reticulum is responsible for hypersensitivity to caffeine and halothane in malignant hyperthermia.
Malignant hyperthermia (MH) is an autosomal dominant myopathy. Molecular genetic studies have shown that the alteration of Arg615 to Cys in the Ca2+ release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor) is cosegregated with porcine MH (Fujii, J., Otsu, K., Zorzato, F., de Leon, S., Khanna, V. K., Weiler, J. E., O'Brien, P. J., and MacLennan, D. H. (1991) Science 253, 448-451; Otsu, K., Khanna, V. K., Archibald, A., and MacLennan, D. H. (1991) Genomics 11, 744-750). Here, using the fluorescence calcium indicator indo-1, we determined the concentration of ionized cytosolic calcium in myoblastic cells transfected with either the wild-type or mutated ryanodine receptor cDNA. The cells expressing the mutant ryanodine receptor showed higher sensitivity to caffeine, which induces Ca2+ release from the sarcoplasmic reticulum through the ryanodine receptor. Exposure to clinical doses of halothane resulted in a rapid increase of [Ca2+]i in cells expressing the mutated ryanodine receptor, whereas no [Ca2+] changes were observed in cells expressing the wild-type ryanodine receptor. These results provide definite evidence that a single amino acid mutation, Arg615-->Cys, in the ryanodine receptor is causative of MH.
Otsu
K
K
First Department of Medicine, Osaka University Medical School, Japan.
Nishida
K
K
Kimura
Y
Y
Kuzuya
T
T
Hori
M
M
Kamada
T
T
Tada
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine; D015153:Blotting, Western; D002110:Caffeine / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D003545:Cysteine; D004342:Drug Hypersensitivity / Q000235:genetics*; D015723:Gene Library; D006221:Halothane / Q000633:toxicity*; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D008305:Malignant Hyperthermia / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008861:Microsomes / Q000378:metabolism; D009124:Muscle Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism*; D017354:Point Mutation; D011061:Poly A / Q000302:isolation & purification / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger; D011817:Rabbits; D019837:Ryanodine Receptor Calcium Release Channel; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D014162:Transfection
D015220:Calcium Channels; D009124:Muscle Proteins; D012333:RNA, Messenger; D019837:Ryanodine Receptor Calcium Release Channel; D011061:Poly A; D002110:Caffeine; D012313:RNA; D001120:Arginine; D003545:Cysteine; D002118:Calcium; D006221:Halothane