The interaction of an impermeant cation with the sheep cardiac RyR channel alters ryanoid association.
In previous studies, we have demonstrated that the interaction of ryanoids with the sarcoplasmic reticulum Ca(2+)-release channel [ryanodine receptor (RyR)] incorporated into planar lipid bilayers reduced the effectiveness of tetraethylammonium (TEA(+)) as a blocker of K(+) translocation (J Gen Physiol 117: 385-393, 2001). In the current study, we investigated both the effect of TEA(+) on [(3)H]ryanodine binding and the actions of this impermeant cation on the interaction of the reversible ryanoid 21-amino-9alpha-hydroxyryanodine with individual, voltage-clamped RyR channels. A dose-dependent inhibition of [(3)H]ryanodine binding was observed in the presence of TEA(+), suggesting that the cation and alkaloid compete for access to a common site of interaction. Single channel studies gave further insights into the mechanism of the competition between the two classes of ligands. TEA(+) decreases the association rate of 21-amino-9alpha-hydroxyryanodine with its receptor, whereas the dissociation rate of the ryanoid from the channel was unaffected. Our results demonstrate that TEA(+) inhibits both K(+) translocation through RyR, and ryanoid interaction at the high affinity ryanodine site on the channel. These actions involve binding of TEA(+) to different, but weakly interacting, sites in the RyR channel.
Cardiac Medicine, National Heart and Lung Institute, Imperial College London, Guy Scadding Building, Dovehouse Street, London SW3 6LY, UK.
Bhavna
B
Tanna
William
W
Welch
Luc
L
Ruest
John L
JL
Sutko
Alan J
AJ
Williams
D000818:Animals; D002414:Cations, Monovalent / Q000378:metabolism; D017136:Ion Transport / Q000187:drug effects; D008024:Ligands; D011188:Potassium / Q000378:metabolism; D026902:Potassium Channel Blockers / Q000494:pharmacology*; D012433:Ryanodine / Q000031:analogs & derivatives / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects* / Q000378:metabolism*; D012756:Sheep; D019789:Tetraethylammonium / Q000494:pharmacology*
C113821:21-amino-9-hydroxyryanodine; D002414:Cations, Monovalent; D008024:Ligands; D026902:Potassium Channel Blockers; D019837:Ryanodine Receptor Calcium Release Channel; D012433:Ryanodine; D019789:Tetraethylammonium; D011188:Potassium