TY - JOUR T1 - Pharmacological and Histochemical Distinctions Between Molecularly Defined Sarcolemmal K<sub>ATP</sub> Channels and Native Cardiac Mitochondrial K<sub>ATP</sub> Channels JF - Molecular Pharmacology JO - Mol Pharmacol SP - 1000 LP - 1005 DO - 10.1124/mol.55.6.1000 VL - 55 IS - 6 AU - Hai Hu AU - Toshiaki Sato AU - Jegatheesan Seharaseyon AU - Yongge Liu AU - David C. Johns AU - Brian O’Rourke AU - Eduardo Marbán Y1 - 1999/06/01 UR - http://molpharm.aspetjournals.org/content/55/6/1000.abstract N2 - A variety of direct and indirect techniques have revealed the existence of ATP-sensitive potassium (KATP) channels in the inner membranes of mitochondria. The molecular identity of these mitochondrial KATP (mitoKATP) channels remains unclear. We used a pharmacological approach to distinguish mitoKATP channels from classical, molecularly defined cardiac sarcolemmal KATP (surfaceKATP) channels encoded by the sulfonylurea receptor SUR2A and the pore-forming subunit Kir6.2. SUR2A and Kir6.2 were expressed in human embryonic kidney (HEK)293 cells, and their activities were measured by patch-clamp recordings of membrane current. SurfaceKATP channels are activated potently by 100 μM pinacidil but only weakly by 100 μM diazoxide; in addition, they are blocked by 10 μM glibenclamide, but are insensitive to 500 μM 5-hydroxydecanoate. This pharmacology, which was confirmed with patch-clamp recordings in intact rabbit ventricular myocytes, contrasts with that of mitoKATP channels as indexed by flavoprotein oxidation. MitoKATP channels in myocytes are activated equally by 100 μM diazoxide and 100 μM pinacidil. In contrast to its lack of effect on surfaceKATP channels, 5-hydroxydecanoate is an effective blocker of mitoKATPchannels. Glibenclamide’s effects on mitoKATP channels are difficult to assess, because it independently activates flavoprotein fluorescence, consistent with a previously described primary uncoupling effect. Confocal imaging of the subcellular distribution of expressed fluorescent Kir6.2 in HEK cells and in myocytes revealed no targeting of mitochondrial membranes. The differences in drug sensitivity and subcellular localization indicate that mitoKATP channels are distinct from surface KATP channels at a molecular level. ER -