Ryanodine receptor type 1 (RyR1) possessing malignant hyperthermia mutation R615C exhibits heightened sensitivity to dysregulation by non-coplanar 2,2',3,5',6-pentachlorobiphenyl (PCB 95).
Malignant hyperthermia (MH) susceptibility is conferred by inheriting one of >60 missense mutations within the highly regulated microsomal Ca(2+) channel known as ryanodine receptor type 1 (RyR1). Although MH susceptible patients lack overt clinical signs, a potentially lethal MH syndrome can be triggered by exposure to halogenated alkane anesthetics. This study compares how non-coplanar 2,2',3,5',6-pentachlorobiphenyl (PCB 95), a congener identified in environmental and human samples, alters the binding properties of [(3)H]ryanodine to RyR1 in vitro. Junctional sarcoplasmic reticulum (SR) was isolated from skeletal muscle dissected from wild type pigs ((Wt)RyR1) and pigs homozygous for MH mutation R615C ((MH)RyR1), a mutation also found in humans. Although the level of (Wt)RyR1 and (MH)RyR1 expression is the same, (MH)RyR1 shows heightened sensitivity to activation and altered regulation by physiological cations. We report here that (MH)RyR1 shows more pronounced activation by Ca(2+), and is less sensitive to channel inhibition by Ca(2+) and Mg(2+), compared to (Wt)RyR1. In a buffer containing 100nM free Ca(2+), conditions typically found in resting cells, PCB 95 (50-1000nM) enhances the activity of (MH)RyR1 whereas it has no detectable effect on (Wt)RyR1. PCB 95 (2microM) decreases channel inhibition by Mg(2+) to a greater extent in (MH)RyR1 (IC(50) increased nine-fold) compared to (Wt)RyR1 (IC(50) increased by 2.5-fold). PCB95 reduces inhibition by Ca(2+) two-fold more with (MH)RyR1 than (Wt)RyR1. Our data suggest that non-coplanar PCBs are more potent and efficacious toward (MH)RyR1 than (Wt)RyR1, and have more profound effects on its cation regulation. Considering the important roles of Ca(2+) and Mg(2+) in regulating Ca(2+) signals involving RyR channels, these data provide the first mechanistic evidence that a genetic mutation known to confer susceptibility to pharmacological agents also enhances sensitivity to an environmental contaminant.
Ta
Tram Anh
TA
UC Davis, Center for Children's Environmental Health, Davis, CA 95616, USA.
Pessah
Isaac N
IN
D000818:Animals; D001120:Arginine / Q000235:genetics*; D002118:Calcium / Q000378:metabolism; D003545:Cysteine / Q000235:genetics*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D066298:In Vitro Techniques; D020128:Inhibitory Concentration 50; D008305:Malignant Hyperthermia / Q000235:genetics* / Q000473:pathology; D018482:Muscle, Skeletal / Q000187:drug effects / Q000648:ultrastructure; D009154:Mutation; D011078:Polychlorinated Biphenyls / Q000633:toxicity*; D011485:Protein Binding / Q000187:drug effects; D012433:Ryanodine / Q000493:pharmacokinetics; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics* / Q000502:physiology; D012519:Sarcoplasmic Reticulum / Q000187:drug effects; D013552:Swine; D014316:Tritium / Q000493:pharmacokinetics
D019837:Ryanodine Receptor Calcium Release Channel; C032904:2,2',3,5',6-pentachlorobiphenyl; D014316:Tritium; D012433:Ryanodine; D001120:Arginine; D011078:Polychlorinated Biphenyls; D003545:Cysteine; D002118:Calcium