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ATP-sensitive potassium channels in adult mouse skeletal muscle: Characterization of the ATP-binding site

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Summary

Single K+-selective channels were studied in excised inside-out membrane patches from dissociated mouse toe muscle fibers. Channels of 74 pS conductance in symmetrical 160mm KCl solutions were blocked reversibly by 10 μm internal ATP and thus identified as ATP-sensitive K+ channels. The channels were also blocked reversibly bymm concentrations of internal adenosine, adenine and thymine, but not by cytosine and uracil. The efficacy of the reversible channel blockers was higher when they were present in internal NaCl instead of KCl solutions. An irreversible inhibition of ATP-sensitive K+ channels was observed after application of several sulphydryl-modifying substances in the internal solution: 0.5mm chloramine-T, 50mm hydrogen peroxide or 2mm n-ethylmaleimide (NEM). Largeconductance Ca-activated K+ channels were not affected by these reagents. The presence of 1mm internal ATP prevents the irreversible inhibition of ATP-sensitive K+ channels by NEM. The results suggest that internal Na+ ions increase the affinity of the ATP-sensitive K+ channel to ATP and to other reversible channel blockers and that a functionally important SH-group is located at or near the ATP-binding site.

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References

  • Allen, C.N., Akaike, A., Albuquerque, E.X. 1984. The frog interosseal muscle fiber as a new model for patch clamp studies of chemosensitive- and voltage-sensitive ion channels: Actions of acetylcholine and batrachotoxin.J. Physiol. (Paris) 79:338–343

    Google Scholar 

  • Ashcroft, F.M. 1988. Adenosine 5′-triphosphate-sensitive potassium channels.Annu. Rev. Neurosci. 11:97–118

    Article  PubMed  Google Scholar 

  • Ashcroft, F.M., Kakei, M. 1987. Effects of internal Mg2+ on ATP-sensitive K-channels in isolated rat pancreatic B-cells.J. Physiol. (London) 390:72P

    Google Scholar 

  • Ashford, M.L.J., Sturgess, N.C., Trout, N.J., Gardner, N.J., Hales, C.N. 1988. Adenosine-5′-triphosphate-sensitive ion channels in neonatal rat cultured central neurones.Pfluegers Arch. 412:297–304

    Article  Google Scholar 

  • Brehm, P., Kullberg, R. 1987. Acetylcholine receptor channels on adult mouse skeletal muscle are functionally identical in synaptic and nonsynaptic membrane.Proc. Natl. Acad. Sci. USA 84:2550–2554

    PubMed  Google Scholar 

  • Burton, F., Dörstelmann, U., Hutter, O.F. 1988. Single-channel activity in sarcolemmal vesicles from human and other mammalian muscles.Muscle Nerve 11:1029–1038

    Article  PubMed  Google Scholar 

  • Colquhoun, D., Sigworth, F.J. 1983. Fitting and statistical analysis of single-channel records.In: Single-Channel Recordings. B. Sakmann and E. Neher, editors. pp. 191–263. Plenum, New York

    Google Scholar 

  • Cook, D.L., Hales, N. 1984. Intracellular ATP directly blocks K+ channels in pancreatic B-cells.Nature (London) 311:271–273

    Article  Google Scholar 

  • Dunne, M.J., Findlay, I., Petersen, O.H. 1988. Effects of pyridine nucleotides on the gating of ATP-sensitive potassium channels in insulin-secreting cells.J. Membrane Biol. 102:205–216

    Google Scholar 

  • Dunne, M.J., Findlay, I., Petersen, O.H., Wollheim, C.B. 1986. ATP-sensitive K+ channels in an insulin-secreting cell line are inhibited byd-glyceraldehyde and activated by membrane permeabilization.J. Membrane Biol. 93:271–279

    Google Scholar 

  • Dunne, M.J., Illot, M.C., Petersen, O.H. 1987. Interaction of diazoxide, tolbutamine and ATP4− on nucleotide-dependent K+ channels in an insulin-secreting cell line.J. Membrane Biol. 99:215–224

    Google Scholar 

  • Findlay, I. 1988. ATP4− and ATP·Mg inhibit the ATP-sensitive K+ channel of rat ventricular myocytes.Pfluegers Arch. 412:37–41

    Google Scholar 

  • Flügge, U.-I., Heldt, H.W. 1977. Specific labelling of a protein involved in phosphate transport of chloroplasts by pyridoxal-5′-phosphate.FEBS Lett. 82:29–33

    Article  PubMed  Google Scholar 

  • 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

    Article  Google Scholar 

  • Hegyvary, C., Post, R.L. 1971. Binding of adenosine triphosphate to sodium and potassium ion-stimulated adenosine triphosphatase.J. Biol. Chem. 246:5234–5240

    PubMed  Google Scholar 

  • Horie, M., Irisawa, H., Noma, A. 1987. Voltage-dependent magnesium block of adenosine-triphosphate-sensitive potassium channel in guinea-pig ventricular cells.J. Physiol. (London) 387:251–272

    Google Scholar 

  • Kakei, M., Noma, A., Shibasaki, T. 1985. Properties of adenosine-triphosphate-regulated potassium channels in guinea-pig ventricular cells.J. Physiol. (London) 363:441–462

    Google Scholar 

  • Kumazawa, N., Tusjimoto, T., Fukushima, Y. 1986. Influence of voltage and ATP on ion-channel of (Na,K)ATPase incorporated into solvent-free phospholipid planar bilayers.Biochem. Biophys. Res. Commun. 136:767–772

    Article  PubMed  Google Scholar 

  • Lundblad, R.L. 1984. Chemical reagents for protein modification. Vol. 1: Proteins. CRC, Boca Raton, Florida

    Google Scholar 

  • Noma, A. 1983. ATP-regulated K+ channels in cardiac muscle.Nature (London) 305:147–148

    Article  Google Scholar 

  • Ohno-Shosaku, T., Zünkler, B.J., Trube, G. 1987. Dual effects of ATP on K+ currents of mouse pancreatic β-cells.Pfluegers Arch. 408:133–138

    Article  Google Scholar 

  • Patzelt-Wenczler, R., Pauls, H., Erdmann, E., Schoner, W. 1975. Evidence for a sulfhydryl group in the ATP-binding site of (Na++K+)-activated ATPase.Eur. J. Biochem. 53:301–311

    Article  Google Scholar 

  • Quayle, J.M., Stanfield, P.R. 1989. Block of the ATP-sensitive potassium channel of frog skeletal muscle by internal sodium ions.J. Physiol. (London) 410:85P (Abstr.)

    Google Scholar 

  • Rack, M., Hu, S.-L., Rubly, N., Waschow, C. 1984. Effects of chemical modification of amino and sulfhydryl groups on the voltage-clamped frog node of Ranvier.Pfluegers Arch. 400:403–408

    Article  Google Scholar 

  • Ribalet, B., Ciani, S. 1987. Regulation by cell metabolism and adenine nucleotides of a K channel in insulin-secreting B-cells (RINm5F).Proc. Natl. Acad. Sci. USA 84:1721–1725

    PubMed  Google Scholar 

  • Salem, N., Jr., Lauter, C.J., Trams, E.G. 1981. Selective chemical modification of plasma membrane ectoenzymes.Biochim. Biophys. Acta 641:366–376

    PubMed  Google Scholar 

  • Schuurmans Stekhoven, F., Bonting, S.L. 1981. Transport adenosine triphosphatases: properties and functions.Physiol Rev. 61:1–76

    PubMed  Google Scholar 

  • Skou, J.C. 1974a. Effect of ATP on the intermediary steps of the reaction of the (Na++K+)-dependent enzyme system: I. Studied by the use ofn-ethylmaleimide inhibition as a tool.Biochim. Biophys. Acta 339:234–245

    Google Scholar 

  • Skou, J.C. 1974b. Effect of ATP on the intermediary steps of the reaction of the (Na++K+)-dependent enzyme systems: II. Effect of a variation in the ATP/Mg2+ ratio.Biochim. Biophys. Acta 339:246–257

    Google Scholar 

  • Smyth, D.G., Blumenfeld, O.O., Konigsberg, W. 1964. Reactions ofn-ethylmaleimide with peptides and amino acids.Biochem. J. 91:589–595

    PubMed  Google Scholar 

  • Spruce, A.E., Standen, N.B., Stanfield, P.R. 1985. Voltage-dependent ATP-sensitive potassium channels of skeletal muscle membrane.Nature (London) 316:736–738

    Article  Google Scholar 

  • Spruce, A.E., Standen, N.B., Stanfield, P.R. 1987. Studies of the unitary properties of adenosine-5′-triphosphate-regulated potassium channels of frog skeletal muscle.J. Physiol. (London) 382:213–236

    Google Scholar 

  • Weik, R., Neumcke, B. 1988. A sulfhydryl group on the ATP-sensitive K channel in mouse skeletal muscle.Pfluegers Arch. 412 (Suppl. No. 1):R11 (Abstr.)

    Google Scholar 

  • Woll, K.H., Leibowitz, M.D., Neumcke, B., Hille, B. 1987. A high-conductance anion channel in adult amphibian skeletal muscle.Pfluegers Arch. 410:632–640

    Google Scholar 

  • Zilberter, Y., Burnashev, N., Papin, A., Portnov, V., Khodorov, B. 1988. Gating kinetics of ATP-sensitive single potassium channels in myocardial cells depends on electromotive force.Pfluegers Arch. 411:584–589

    Google Scholar 

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Weik, R., neumcke, B. ATP-sensitive potassium channels in adult mouse skeletal muscle: Characterization of the ATP-binding site. J. Membrain Biol. 110, 217–226 (1989). https://doi.org/10.1007/BF01869152

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