HMG CoA reductase inhibitor-induced myotoxicity: pravastatin and lovastatin inhibit the geranylgeranylation of low-molecular-weight proteins in neonatal rat muscle cell culture.
In previous studies, inhibition of cholesterol synthesis by HMG CoA reductase inhibitors (HMGRI) was associated with myotoxicity in cultures of neonatal rat skeletal myotubes, and rhabdomyolysis in rats, rabbits, and humans in vivo. In vitro myotoxicity was directly related to HMGRI-induced depletion of mevalonate, farnesol, and geranylgeraniol, since supplementation with these intermediate metabolites abrogated the toxicity. Both farnesol and geranylgeraniol are required for the posttranslational modification, or isoprenylation, of essential regulatory proteins in mammalian cells. The objective of the present study was to measure changes in protein isoprenylation in cultured neonatal rat skeletal muscle cells exposed for 24 hr to increasing concentrations of pravastatin or lovastatin. Proteins were labeled with [3H]mevalonate, [3H]farnesyl pyrophosphate (FPP), or [3H]geranylgeranyl pyrophosphate (GGPP), and then separated by SDS-PAGE and quantitated by scintillation counting and densitometry of autoradiographs. Mevalonate and FPP labeling of the majority of proteins increased in a concentration-dependent manner, even at concentrations greater than 2 microM lovastatin and 25 microM pravastatin that completely inhibited cholesterol synthesis. In contrast, mevalonate and FPP labeling of three protein bands with molecular weights of 26.6, 27.7, and 28.9 kDa was markedly inhibited at concentrations higher than 1 microM lovastatin and 400 microM pravastatin, which inhibited protein synthesis and disrupted myotube morphology after longer exposures in a previous study. In contrast, these proteins were equally well labeled by GGPP at all HMGRI concentrations tested, suggesting that isoprenylation of the 26.9-, 27.8-, and 28.9-kDa proteins requires geranylgeraniol. The results of this study indicate that HMGRI-induced myotoxicity is most likely related to reduced posttranslational modification of specific regulatory proteins by geranylgeraniol.
Flint
O P
OP
Department of Experimental Pathology, Bristol-Myers Squibb Pharmaceutical Research Institute, Syracuse, New York 13057, USA. invitro@invitro.org
Masters
B A
BA
Gregg
R E
RE
Durham
S K
SK
D000818:Animals; D000831:Animals, Newborn; D000924:Anticholesteremic Agents / Q000633:toxicity*; D001345:Autoradiography; D002478:Cells, Cultured; D002784:Cholesterol / Q000096:biosynthesis; D003720:Densitometry; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D007553:Isotope Labeling; D008148:Lovastatin / Q000633:toxicity*; D008798:Mevalonic Acid / Q000378:metabolism; D008970:Molecular Weight; D009124:Muscle Proteins / Q000187:drug effects / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000378:metabolism; D017035:Pravastatin / Q000633:toxicity*; D011247:Pregnancy; D011499:Protein Processing, Post-Translational / Q000187:drug effects / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012588:Scintillation Counting; D012717:Sesquiterpenes; D014316:Tritium
D000924:Anticholesteremic Agents; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D009124:Muscle Proteins; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; D014316:Tritium; C004808:farnesyl pyrophosphate; D002784:Cholesterol; D008148:Lovastatin; D017035:Pravastatin; C002963:geranylgeranyl pyrophosphate; D008798:Mevalonic Acid