The effect of intracellular anions on ATP-dependent potassium channels of rat skeletal muscle.
1. We have used excised inside-out patches to study the effects of anions bathing the cytoplasmic surface of the membrane on ATP-dependent K+ channels of rat flexor digitorum brevis muscle. Channels were closed by ATP applied to the cytoplasmic face of the patch with a concentration for half-closure (Ki) of 14 microM, were highly selective for K+ and had unitary conductances of 62 pS in symmetrical 155 mM K+ and 27 pS in 5 mM [K+]o. 2. In 139 mM Cl- internal solution channel activity declined rapidly after excision of the patch. Inclusion of 40 mM potassium gluconate (substituted for KCl) in the solution both restored channel activity and greatly slowed its subsequent run-down. 3. The action of gluconate was concentration dependent. The effect did not involve a change in ATP binding, since the Ki for ATP was not significantly changed by gluconate, and was specific for the cytoplasmic face of the patch. 4. The anions pyruvate, lactate and acetate were all able to restore channel activity after run-down, though less well than gluconate, while sulphate and methylsulphate were without effect. 5. Analysis of single channel kinetics showed that gluconate did not affect mean open lifetime, but led to a decrease in the number and duration of long closings. 6. Anions are most likely to act by stabilizing the structure of the channel protein. Changes in the intracellular concentration of certain anions may play a role in regulating channel activity.
Department of Cell Physiology and Pharmacology, University of Leicester.
H C
HC
McKillen
N W
NW
Davies
P R
PR
Stanfield
N B
NB
Standen
D000255:Adenosine Triphosphate / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D000838:Anions / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism; D005942:Gluconates / Q000378:metabolism; D007700:Kinetics; D018482:Muscle, Skeletal / Q000294:innervation / Q000378:metabolism*; D009431:Neural Conduction / Q000502:physiology; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D000838:Anions; D005942:Gluconates; D015221:Potassium Channels; D000255:Adenosine Triphosphate