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The effects of the covalent modifier of amino groups, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) on the single-channel properties of purified sheep cardiac ryanodine receptors (RyR) incorporated into planar phospholipid bilayers were investigated. DIDS increased single-channel conductance and open probability (P(o)) and induced unique modifications to the voltage-dependence of gating. The effects of DIDS on conduction and gating were irreversible within the time scale of the experiments, and both effects were dependent on the permeant ion. DIDS induced a greater increase in conductance with Ca(2+) (20%) compared with K(+) (8%) as the permeant ion. After modification by DIDS, all channels could be rapidly inactivated in a voltage-dependent manner. The open probability of the DIDS-modified channel decreased with increasing positive or negative transmembrane potentials; however, inactivation was only observed at negative potentials. Our results demonstrate that inactivation of RyR channels is dependent on the ligand activating the channel, and this will have consequences for the control and termination of sarcoplasmic reticulum Ca(2+) release in cardiac cells.

Original publication

DOI

10.1016/S0006-3495(02)75644-0

Type

Journal

Biophys J

Publication Date

06/2002

Volume

82

Pages

3037 - 3047

Keywords

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid, Animals, Biophysical Phenomena, Biophysics, Calcium, Electric Conductivity, In Vitro Techniques, Ion Channel Gating, Kinetics, Ligands, Membrane Potentials, Models, Biological, Myocardium, Potassium, Ryanodine Receptor Calcium Release Channel, Sheep