Inhibition of the mitochondrial KATP channel by long-chain acyl-CoA esters and activation by guanine nucleotides.
The mitochondrial KATP channel (mitoKATP) is highly sensitive to ATP, which inhibits K+ flux with K1/2 values of 20-40 microM. This raises the question, how can mitoKATP be opened in the presence of physiological concentrations of ATP? We measured K+ flux in liposomes reconstituted with purified mitoKATP and found that guanine nucleotides are potent activators of this channel. ATP-inhibited K+ flux was completely reactivated by both GTP (K1/2 = 7 microM) and GDP (K1/2 = 140 microM). These ligands had no effect in the absence of ATP. The K1/2 for ATP inhibition exhibited quadratic dependence on [GTP] and [GDP], consistent with two binding sites for guanine nucleotides. We also found that palmitoyl-CoA and oleoyl-CoA inhibited K+ flux through reconstituted mitoKATP with K1/2 values of 260 nM and 80 nM, respectively. This inhibition was reversed by GTP (K1/2 = 232 microM) as well as by the K+ channel openers cromakalim (20 microM) and diazoxide (10 microM). Inhibition of mitoKATP by long-chain acyl-CoA esters, like that of ATP, exhibited an absolute requirement for Mg2+ ions. We propose that the open-closed state of the mitochondrial KATP channel is determined by the relative cytosolic concentrations of GTP and long-chain acyl-CoA esters.
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Department of Chemistry, Biochemistry, and Molecular Biology, Oregon Graduate Institute of Science & Technology, Portland, Oregon 97291-1000, USA. garlid@admin.ogi.edu
Yarov-Yarovoy
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D000214:Acyl Coenzyme A / Q000494:pharmacology*; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology; D019806:Cromakalim; D003981:Diazoxide / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000494:pharmacology*; D006160:Guanosine Triphosphate / Q000494:pharmacology*; D008274:Magnesium / Q000494:pharmacology; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism; D010171:Palmitoyl Coenzyme A / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism; D026902:Potassium Channel Blockers; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats
D000214:Acyl Coenzyme A; D001578:Benzopyrans; D026902:Potassium Channel Blockers; D011758:Pyrroles; D019806:Cromakalim; D006153:Guanosine Diphosphate; C017585:oleoyl-coenzyme A; D010171:Palmitoyl Coenzyme A; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D008274:Magnesium; D003981:Diazoxide; D011188:Potassium