Ryanoid modification of the cardiac muscle ryanodine receptor channel results in relocation of the tetraethylammonium binding site

The interaction of ryanodine and derivatives of ryanodine with the high aff inity binding site on the ryanodine receptor (RyR) channel brings about a c haracteristic modification of channel function. In all cases, channel open probability increases dramatically and single-channel current amplitude is reduced. The amplitude of the ryanoid-modified conductance state is determi ned by structural features of the ligand. An investigation of ion handing i n the ryanodine-modified conductance state has established that reduced con ductance results from changes in both the affinity of the channel for perme ant ions and the relative permeability of ions within the channel (Lindsay, A.R.G., A. Tinker, and A.J. Williams. 1994. J. Gen. Physiol. 104:425-447). It has been proposed that these alterations result from a reorganization o f channel structure induced by the binding of the ryanoid. The experiments reported here provide direct evidence for ryanoid-induced restructuring of RyR. TEA(+) is a concentration- and voltage-dependent blocker of RyR in the absence of ryanoids. We have investigated block of K+ current by TEA(+) in the unmodified open state and modified conductance states of RyR induced b y 21-amino-9 alpha -hydroxyryanodine, 21-azido-9 alpha -hydroxyryanodine, r yanodol, and 21-p-nitrobenzoylamino-9 alpha -hydroxyryanodine. Analysis of the voltage dependence of block indicates that the interaction of ryanoids with RyR leads to an alteration in this parameter with an apparent relocati on of the TEA(+) blocking site within the voltage drop across the channel a nd an alteration in the affinity of the channel for the blocker. The degree of change of these parameters correlates broadly with the change in conduc tance of permeant cations induced by the ryanoids, indicating that modifica tion of RyR channel structure by ryanoids is likely to underlie both phenom ena.