Rapamycin inhibits activation of ryanodine receptors from skeletal muscle by the fatty acyl CoA-acyl CoA binding protein complex.
We previously showed (Fulceri et al., Biochem. J. 325, 423, 1997) that the fatty acyl CoA ester palmitoyl CoA (PCoA) complexed with a molar excess of its cytosolic binding protein (ACBP) causes a discrete Ca(2+) efflux or allows Ca(2+) release by suboptimal caffeine concentrations, in the Ca(2+)-preloaded terminal cisternae fraction (TC) from rabbit skeletal muscle, by activating ryanodine receptor Ca(2+) release channels (RyRC). We show here that both effects were abolished by pretreating TC with the FKBP12 ligand rapamycin (20 microM). Moreover, rapamycin reversed the Ca(2+) release induced by combined treatment with 3 mM caffeine and the PCoA-ACBP complex. Rapamycin also reduced the Ca(2+)-releasing activity by PCoA alone. Under the above experimental conditions, rapamycin removed FKBP12 from the TC membranes, as revealed by Western blot analysis. We conclude that FKBP12 associated with RyRC in the TC membrane participates in the activation of the Ca(2+) channel by fatty acyl CoA esters.
Fulceri
R
R
Dipartimento di Fisiopatologia e Medicina Sperimentale, Università di Siena, Siena, 53100-, Italy.
Giunti
R
R
Knudsen
J
J
Leuzzi
R
R
Kardon
T
T
Benedetti
A
A
D000818:Animals; D015153:Blotting, Western; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D003600:Cytosol / Q000378:metabolism; D026261:Diazepam Binding Inhibitor; D020104:Immunophilins / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism*; D010171:Palmitoyl Coenzyme A / Q000037:antagonists & inhibitors*; D010957:Plasmids; D011817:Rabbits; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects*; D020123:Sirolimus / Q000494:pharmacology*; D022021:Tacrolimus Binding Proteins
D002352:Carrier Proteins; D026261:Diazepam Binding Inhibitor; D007166:Immunosuppressive Agents; D019837:Ryanodine Receptor Calcium Release Channel; D010171:Palmitoyl Coenzyme A; D002110:Caffeine; D022021:Tacrolimus Binding Proteins; D020104:Immunophilins; D002118:Calcium; D020123:Sirolimus