AMP is a partial agonist at the sheep cardiac ryanodine receptor.
We have investigated the ability of AMP to modulate the native sheep cardiac ryanodine receptor (RyR) channel at various cytosolic [Ca2+]. Channels were incorporated into planar phospholipid bilayers and current fluctuations through the bilayer were monitored under voltage clamp conditions. We demonstrate that AMP only exhibits agonist activity if the cytosolic [Ca2+] is sufficiently high. Even in the presence of a high cytosolic [Ca2+] (65 microM), AMP cannot fully open the channel and the maximum open probability (Po) observed is approximately 0.3 at 2 mM AMP. Concentrations of AMP above the maximally activating level cause inactivation of the channel. Our experiments indicate that AMP is an agonist with such low efficacy at the ATP sites on the cardiac RyR that it is effectively an antagonist of ATP-induced increases in Po. Our study demonstrates that the number of phosphates attached to the 5'-carbon of the ribose ring of adenine-based compounds determines the efficacy of the ligand to increase the Po of the cardiac RyR. Substitution of groups at this position may lead to the identification of potent antagonists at ATP sites on RyR.
Ching
L L
LL
Cardiac Medicine, Imperial College School of Medicine, London.
Williams
A J
AJ
Sitsapesan
R
R
D000249:Adenosine Monophosphate / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000465:Algorithms; D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000502:physiology; D002120:Calcium Channel Agonists / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D006321:Heart / Q000187:drug effects*; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects / Q000378:metabolism*; D012756:Sheep
D002120:Calcium Channel Agonists; D019837:Ryanodine Receptor Calcium Release Channel; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D002118:Calcium