Abstract
Elementary K+ currents were recorded at 19°C in inside-out patches from cultured neonatal rat cardiocytes to elucidate the block phenomenology in cardiac ATP-sensitive K+ channels when inhibitory drug molecules, such as the sulfonylurea glibenclamide, the phenylalkylamine verapamil or sulfonamide derivatives (HE 93 and sotalol), are interacting in an attempt to stress the hypothesis of multiple channel-associated drug targets.
Similar to their adult relatives, neonatal cardiac K(ATP) channels are characterized by very individual open state kinetics, even in cytoplasmically well-controlled, cell-free conditions; at −7 mV, τopen(1) ranged from 0.7 to 4.9 msec in more than 200 patches and τopen(2) from 10 to 64 msec—an argument for a heterogeneous channel population. Nevertheless, a common response to drugs was observed. Glibenclamide and the other inhibitory molecules caused long-lasting interruptions of channel activity, after cytoplasmic application, as if drug occupancy trapped cardiac K(ATP) channels in a very stable, nonconducting configuration. The resultant NP 0 depression was strongest with glibenclamide (apparent IC50 13 nmol/liter) and much weaker with verapamil (apparent IC50 9 μmol/liter), HE 93 (apparent IC50 29 μmol/liter) and sotalol (apparent IC50 43 μmol/ liter) and may have resulted from the occupancy of a single site with drug-specific affinity or of two sites, the high affinity glibenclamide target and a distinct nonglibenclamide, low affinity target.
Changes in open state kinetics, particularly in the transition between the O1 state and the O2 state, are other manifestations of drug occupancy of the channel. Any inhibitory drug molecule reduced the likelihood of attaining the O2 state, consistent with a critical reduction of the forward rate constant governing the O1-O1 transition. But only HE 93 (10 μmol/liter) associated (with an apparent association rate constant of 2.3 × 106 mol−1 sec−1) to shorten significantly τopen(2) to 60.6 ± 6% of the predrug value, not the expected result when the entrance in and the exit from the O2 state would be drug-unspecifically nfluenced. Sotalol found yet another and definitely distinctly located binding site to interfere with K+ permeation; both enantiomers associated with a rate close to 5×105 mol−1 sec−1 with the open pore thereby flicker-blocking cardiac K(ATP) channels. Clearly, these channels accommodate more than one drug-binding domain.
Similar content being viewed by others
References
Argentieri, T.M., Troy, H.H., Carroll, M.S., Doroshuk, C.M., Sullivan, M.E. 1993. Electrophysiologic activity and antiarrhythmic efficacy of CK-3579, a new class III antiarrhythmic agent with β-adrenergic properties. J. Cardiovasc. Pharmacol. 21:647–655
Ashcroft, F.M. 1988. Adenosine 5′-triphosphate-sensitive potassium channels. Annu. Rev. Neurosci. 11:97–118
Ashcroft, F.M., Ashcroft, S.J.H., Berggren, P.O., Betzholz, C., Rorsman, P., Trube, G. 1988. Expression of K channels in Xenopus laevis oocytes injected with poly(A+)mRNA from the insulinsecreting β-cell line, HIT T15. FEBS Lett. 239:185–189
Benndorf, K., Friedrich, M., Hirche, H.J. 1991. Anoxia opens ATP regulated K channels in isolated heart cells. Pfluegers Arch. 419:108–110
Benz, I., Kohlhardt, M. 1994a. Blockade of cardiac outwardly rectifying K+ channels by TEA and class III antiarrhythmics—evidence against a single drug-sensitive channel site. Eur. Biophys. J. 22:437–446
Benz, I., Kohlhardt, M. 1994b. Chemically modified cardiac Na+ channels and their sensitivity to antiarrhythmics: Is there a hidden drug receptor? J. Membrane Biol. 139:191–201
Blatz, A.L., Magleby, K.L. 1984. Ion conductance and selectivity of single calcium-activated potassium channels in cultured rat muscle. J. Gen. Physiol. 84:1–23
Catterall, W.A., Striessnig, J. 1992. Receptor sites for Ca2+ channel antagonists. TIPS 13:256–262
Colatsky, T.J. 1982. Mechanisms of action of lidocaine and quinidine on action potential duration in rabbit cardiac Purkinje fibers. Circ. Res. 50:17–27
Colquhoun, D., Sigworth, F. 1983. Fitting and statistical analysis of single channel records. In: Single-Channel Recordings. B. Sakmann, and E. Neher, editors, pp. 191–264. Plenum, New York
Cook, D.L., Hales, C.N. 1984. Intracellular ATP directly blocks K+ channels in pancreatic β-cells. Nature 311:271–273
Cook, D.L., Satin, L.S., Ashford, M.L.J., Hales, C.N. 1988. ATP-sensitive K+ channels in pancreatic β-cells. Diabetes 37:495–498
Edwards, G., Weston, A.H. 1993. The pharmacology of ATP-sensitive potassium channels. Annu. Rev. Pharmacol. Toxicol. 33:597–637
Faivre, J.F., Findlay, I. 1990. Action potential duration and activation of ATP-sensitive potassium current in isolated guinea-pig ventricular myocytes. Biochim Biophys Acta 1029:167–172
Findlay, I. 1992. Effects of pH upon the inhibition by sulphonylurea drugs of ATP-sensitive K+ channels in cardiac muscle. J. Pharmacol. Exp. Ther. 262:71–79
Findlay, I., Faivre, J.F. 1991. ATP-sensitive K channels in heart muscle. Spare channel. FEBS Lett. 279:95–97
Fosset, M., deWeille, J.R., Green, R.D., Schmid-Antomarchi, H., Lazdunski, M. 1988. Antidiabetic sulfonylureas control action potential properties in heart cells via high-affinity receptors that are linked to ATP-dependent K+ channels. J. Biol. Chem. 263:7933–7936
French, J.F., Riera, L.C., Mullins, U.L., Sarmiento, J.G. 1991. Modulation of H3 glibenclamide binding to cardiac and insulinoma membranes. Eur. J. Pharmacol. 207:23–28
Gillis, K.D., Gee, W.M., Hammoud, A., McDaniel, M.L., Falke, L.C., Misler, S. 1989. Effects of sulfonamides on a metabolite-regulated ATP-sensitive K+ channel in rat pancreatic B-cells. Am. J. Physiol. 257:C1119-C1127
Gopalakrishnan, M., Johnson, D.E., Janis, R.A., Triggle, D.J. 1991. Characterization and binding of the ATP-sensitive potassium channel ligand, H3 glyburide, to neuronal and muscle preparations. J. Pharmacol. Exp. Ther. 257:1162–1171
Hamill, O.P., Marty, A., Neher, E., Sakmann, B., Sigworth, F.J. 1981. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pfluegers Arch. 391:85–100
Hartmann, H.A., Kirsch, G.E., Drewe, J.A., Taglialatela, M., Joho, R.H., Brown, A.M. 1991. Exchange of conduction pathways between two related K+ channels. Science 251:942–944
Ho, K., Nichols, C.G., Lederer, W.J., Lytton, J., Vassilev, P.M., Kanazirska, M.V., Hebert, S.C. 1993. Cloning and expression of an inwardly rectifying ATP-regulated potassium channel. Nature 362:31–37
Kakei, M., Noma, A. 1984. Adenosine-5′-triphosphate-sensitive single potassium channels in atrioventricular node cells of the rabbit heart. J. Physiol. 352:265–284
Kimura, S., Bassett, A.L., Xi, H., Myerburg, R.J. 1992. Verapamil diminishes action potential changes during metabolic inhibition by blocking ATP-regulated potassium currents. Circ. Res. 71:87–95
Kirsch, G.E., Codina, J., Birnbaumer, L., Brown, A.M. 1990. Coupling of ATP-sensitive K+ channels to A1 receptors by G proteins in rat ventricular myocytes. Am. J. Physiol. 259:H820-H826
Kirsch, G.E., Taglialatela, M., Brown, A.M. 1991. Internal and external TEA block in single cloned K+ channels. Am. J. Physiol. 261:C583-C590
Kohlhardt, M., Fichtner, H., Fröbe, U. 1989. Gating in iodate-modified single cardiac Na+ channels. J. Membrane Biol. 112:67–78
Kubo, Y., Baldwin, T.J., Jan, Y.N., Jan, L.Y. 1993. Primary structure and functional expression of a mouse inward rectifier potassium channel. Nature 362:127–133
Nichols, C.G., Lederer, W.J. 1991. Adenosine triphosphate-sensitive potassium channels in the cardiovascular system. Am. J. Physiol. 261:H1675-H1686
Niki, I., Kelly, R.P., Ashcroft, S.J.H., Ashcroft, F.M. 1989. ATP-sensitive K channels in HIT T15 β-cells studied by patch-clamp methods, 86Rb efflux and glibenclamide binding. Pfluegers Arch. 415:47–55
Niki, I., Nicks, J.L., Ashcroft, S.J.H. 1990. The beat cell glibenclamide receptor is an ADP-binding protein. Biochem. J. 268:713–718
Noma, A. 1983. ATP-regulated K+ channels in cardiac muscle. Nature 305:147–148
Noma, A., Takano, M. 1991. The ATP-sensitive K+ channel. Jap. J. Physiol. 41:177–187
Ohno, T., Zunkler, B.J., Trube, G. 1987. Dual effects of ATP on K+ currents of mouse pancreatic β-cells. Pfluegers Arch. 408:133–138
Panten, U., Burgfeld, J., Goerke, F., Rennicke, M., Schwanstecher, M., Wallasch, A., Zunkler, B.J., Lenzen, S. 1989. Control of insulin secretion by sulfonylureas, meglitinide and diazoxide in relation to their binding to the sulfonylurea receptor in pancreatic islets. Biochem. Pharmacol. 38:1217–1229
Parent, L., Coronado, R. 1989. Reconstitution of the ATP-sensitive potassium channel of skeletal muscle. J. Gen. Physiol. 94:445–463
Ripoll, C., Lederer, W.J., Nichols, C.G. 1990. On the mechanism of inhibition of KATP channels by glibenclamide in rat ventricular cell membranes. Circulation 82:111–12
Spruce, A.E., Standen, N.B., Stanfield, P.R. 1987. Studies on the unitary properties of adenosine-5′-triphosphate-regulated potassium channels of frog skeletal muscle. J. Physiol. 382:213–237
Standen, N.B., Quayle, J.M., Davies, N.W., Brayden, J.E., Huang, Y., Nelsom, M.T. 1989. Hyperpolarizing vasodilators activate ATP-sensitive K+ channels in arterial smooth muscle. Science 245:177–180
Takano, M., Qin, D., Noma, A. 1990. ATP-dependent decay and recovery of K+ channels in guinea pig cardiac myocytes. Am. J. Physiol. 258:H45-H50
Undrovinas, A.I., Burnashev, N., Eroshenko, D., Fleidervish, I., Starmer, C.F., Makielski, J.C., Rosenshtraukh, L.V. 1990. Quinidine blocks adenosine 5′-triphosphate-sensitive potassium channels in heart. Am. J. Physiol. 259:H1609-H1612
Villarroel, A., Alvarez, O., Oberhauser, A., Latorre, R. 1988. Probing a Ca++-activated K+ channel with quaternary ammonium ions. Pfluegers Arch. 413:118–126
Virag, L., Furukawa, T., Hiraoka, M. 1993. Modulation of the effect of glibenclamide on KATP channels by ATP and ADP. Mol. Cell. Biochem. 119:209–215
Vleugels, A., Vereecke, J., Carmeliet, E. 1980. Ionic currents during hypoxia in voltage-clamped cat ventricular muscle. Circ. Res. 47:501–508
Wang, W., Giebisch, G. 1991. Dual modulation of renal ATP-sensitive K+ channels by protein kinases A and C. Proc. Natl. Acad. Sci. USA 88:9722–9725
Weiss, R.E., Horn, R. 1986. Functional differences between two classes of sodium channels in developing rat skeletal muscle. Science 233:361–364
Weiss, J.N., Venkatesh, N., Lamp, S.T. 1992. ATP-sensitive K+ channels and cellular K+ loss in hypoxic and ischemic mammalian ventricle. J. Physiol. 447:649–673
Yellen, G., Jurman, M.E., Abramson, T., McKinnon, R. 1991, Mutations affecting internal TEA blockade identify the probable poreforming region of a K+ channel. Science 251:939–942
Yool, A.J., Schwarz, T.L. 1991. Alteration of ionic selectivity of a K+ channel region by mutation of the H5 region. Nature 349:700–704
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Benz, I., Kohlhardt, M. Distinct modes of blockade in cardiac ATP-sensitive K+ channels suggest multiple targets for inhibitory drug molecules. J. Membarin Biol. 142, 309–322 (1994). https://doi.org/10.1007/BF00233438
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00233438