ATP-sensitive K+ channels from aortic smooth muscle incorporated into planar lipid bilayers.
Glibenclamide binding sites were identified in a membrane preparation from canine aortic smooth muscle. The dissociation constant for [3H]glibenclamide binding was 10 +/- 2 nM, with a density of 420 +/- 108 fmol/mg protein. The properties of ATP-sensitive potassium (KATP) channels from the same membrane preparation incorporated into planar lipid bilayers were investigated. ATP was a potent inhibitor of the channels with half-maximal inhibition of channel activity by 41 microM ATP. Glibenclamide inhibited channel activity, and cromakalim activated the channel in the presence of ATP. Blockers of Ca(2+)-activated K+ (KCa) channels (charybdotoxin and tetraethylammonium ions) did not affect KATP channels in concentrations that caused significant block of KCa channels in bilayers. This membrane preparation should allow further biochemical and functional characterization of KATP channels and glibenclamide receptors in arterial smooth muscle.
Kovacs
R J
RJ
Krannert Institute of Cardiology, Indiana University School of Medicine, Indianapolis 46202.
Nelson
M T
MT
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000378:metabolism*; D001578:Benzopyrans / Q000494:pharmacology; D001665:Binding Sites; D018999:Charybdotoxin; D019806:Cromakalim; D004285:Dogs; D005905:Glyburide / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D015221:Potassium Channels / Q000187:drug effects* / Q000378:metabolism; D011758:Pyrroles / Q000494:pharmacology; D012604:Scorpion Venoms / Q000494:pharmacology; D019789:Tetraethylammonium; D013757:Tetraethylammonium Compounds / Q000494:pharmacology; D014665:Vasodilator Agents / Q000494:pharmacology
D001578:Benzopyrans; D008051:Lipid Bilayers; D015221:Potassium Channels; D011758:Pyrroles; D012604:Scorpion Venoms; D013757:Tetraethylammonium Compounds; D014665:Vasodilator Agents; D019806:Cromakalim; D018999:Charybdotoxin; D019789:Tetraethylammonium; D000255:Adenosine Triphosphate; D005905:Glyburide