Heart mitochondria contain functional ATP-dependent K+ channels.
Recent observations challenged the functional importance or even the existence of mitochondrial ATP-dependent K+ (mitoK(ATP)) channels. In the present study, we determined the presence of K(ATP)-channel subunits in mouse heart mitochondria, and investigated whether known openers or blockers of the channel can alter mitochondrial membrane potential. Investigation of the channel composition was performed with antibodies against K(ATP)-channel subunits, namely the sulfonylurea receptor (SUR1 or SUR2) and the inwardly rectifying K+ channel (Kir6.1 or Kir6.2). Specific Kir6.1 and Kir6.2 proteins were found in the mitochondria by western blotting and immunogold electron microscopy. Neither SUR1 nor SUR2 was present in the mitochondria. In contrast, a mitochondrially enriched low molecular weight SUR2-like band was found at approximately 25 kDa. Mitochondrial-transport tags were identified in the sequences of Kir6.1 and Kir6.2, but not in SUR1 or SUR2. The fluorescent BODIPY-glibenclamide labeling of mitochondria indicated direct sulfonylurea binding. Pharmacological characterization of mitoK(ATP) was performed in isolated respiring heart mitochondria. Fluorescent confocal imaging with the membrane potential-sensitive dye MitoFluorRed showed that glibenclamide application changed membrane potential, while the specific mitoK(ATP)-channel openers, diazoxide or BMS-191095, reversed the effect. Mitochondrially formed peroxynitrite is a physiological opener of the channel. We conclude that a functional K(ATP) channel is present in heart mitochondria, which can be opened by diazoxide or BMS-191095. The channel can be composed of Kir6.1 and Kir6.2 subunits and does not contain either SUR1 or SUR2.
Lacza
Zsombor
Z
Department of Physiology and Pharmacology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA. zlacza@mac.com
Snipes
James A
JA
Miller
Allison W
AW
Szabó
Csaba
C
Grover
Gary
G
Busija
David W
DW
D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D003981:Diazoxide / Q000494:pharmacology; D005905:Glyburide / Q000494:pharmacology; D008564:Membrane Potentials; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D016253:Microscopy, Immunoelectron; D008929:Mitochondria, Heart / Q000378:metabolism* / Q000648:ultrastructure; D009588:Nitrogen Mustard Compounds / Q000494:pharmacology; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism; D040901:Proteomics; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors; D014665:Vasodilator Agents / Q000494:pharmacology
D018528:ATP-Binding Cassette Transporters; C577508:Abcc8 protein, mouse; C090412:BMS 181095; D009588:Nitrogen Mustard Compounds; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors; D014665:Vasodilator Agents; D000255:Adenosine Triphosphate; D003981:Diazoxide; D005905:Glyburide