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. 1996 Feb;70(2):754–761. doi: 10.1016/S0006-3495(96)79615-7

Contributions of a negatively charged residue in the hydrophobic domain of the IRK1 inwardly rectifying K+ channel to K(+)-selective permeation.

E Reuveny 1, Y N Jan 1, L Y Jan 1
PMCID: PMC1224975  PMID: 8789092

Abstract

Inwardly rectifying K+ channels are highly selective for K+ ions and show strong interaction with ions in the pore. Both features are important for the physiological functions of these channels and pose intriguing mechanistic questions of ion permeation. The aspartate residue in the second putative transmembrane segment of the IRK1 inwardly rectifying K+ channel, previously implicated in inward rectification gating due to cytoplasmic Mg2+ and polyamine block, is found in this study to be crucial for the channel's ability to distinguish between K+ and Rb+ ions. Mutation of this residue also perturbs the interaction between the channel pore and the Sr2+ blocking ion. Our studies suggest that this aspartate residue contributes to a selectivity filter near the cytoplasmic end of the pore.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aiyar J., Nguyen A. N., Chandy K. G., Grissmer S. The P-region and S6 of Kv3.1 contribute to the formation of the ion conduction pathway. Biophys J. 1994 Dec;67(6):2261–2264. doi: 10.1016/S0006-3495(94)80710-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biermans G., Vereecke J., Carmeliet E. The mechanism of the inactivation of the inward-rectifying K current during hyperpolarizing steps in guinea-pig ventricular myocytes. Pflugers Arch. 1987 Dec;410(6):604–613. doi: 10.1007/BF00581320. [DOI] [PubMed] [Google Scholar]
  3. Choi K. L., Mossman C., Aubé J., Yellen G. The internal quaternary ammonium receptor site of Shaker potassium channels. Neuron. 1993 Mar;10(3):533–541. doi: 10.1016/0896-6273(93)90340-w. [DOI] [PubMed] [Google Scholar]
  4. Collins A., Somlyo A. V., Hilgemann D. W. The giant cardiac membrane patch method: stimulation of outward Na(+)-Ca2+ exchange current by MgATP. J Physiol. 1992 Aug;454:27–57. doi: 10.1113/jphysiol.1992.sp019253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. De Biasi M., Drewe J. A., Kirsch G. E., Brown A. M. Histidine substitution identifies a surface position and confers Cs+ selectivity on a K+ pore. Biophys J. 1993 Sep;65(3):1235–1242. doi: 10.1016/S0006-3495(93)81154-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fakler B., Brändle U., Glowatzki E., Weidemann S., Zenner H. P., Ruppersberg J. P. Strong voltage-dependent inward rectification of inward rectifier K+ channels is caused by intracellular spermine. Cell. 1995 Jan 13;80(1):149–154. doi: 10.1016/0092-8674(95)90459-x. [DOI] [PubMed] [Google Scholar]
  7. Ficker E., Taglialatela M., Wible B. A., Henley C. M., Brown A. M. Spermine and spermidine as gating molecules for inward rectifier K+ channels. Science. 1994 Nov 11;266(5187):1068–1072. doi: 10.1126/science.7973666. [DOI] [PubMed] [Google Scholar]
  8. Gay L. A., Stanfield P. R. Cs(+) causes a voltage-dependent block of inward K currents in resting skeletal muscle fibres. Nature. 1977 May 12;267(5607):169–170. doi: 10.1038/267169a0. [DOI] [PubMed] [Google Scholar]
  9. Goldman D. E. POTENTIAL, IMPEDANCE, AND RECTIFICATION IN MEMBRANES. J Gen Physiol. 1943 Sep 20;27(1):37–60. doi: 10.1085/jgp.27.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. HODGKIN A. L., HOROWICZ P. The influence of potassium and chloride ions on the membrane potential of single muscle fibres. J Physiol. 1959 Oct;148:127–160. doi: 10.1113/jphysiol.1959.sp006278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. HODGKIN A. L., KATZ B. The effect of sodium ions on the electrical activity of giant axon of the squid. J Physiol. 1949 Mar 1;108(1):37–77. doi: 10.1113/jphysiol.1949.sp004310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hagiwara S., Takahashi K. The anomalous rectification and cation selectivity of the membrane of a starfish egg cell. J Membr Biol. 1974;18(1):61–80. doi: 10.1007/BF01870103. [DOI] [PubMed] [Google Scholar]
  13. Hartmann H. A., Kirsch G. E., Drewe J. A., Taglialatela M., Joho R. H., Brown A. M. Exchange of conduction pathways between two related K+ channels. Science. 1991 Feb 22;251(4996):942–944. doi: 10.1126/science.2000495. [DOI] [PubMed] [Google Scholar]
  14. Harvey R. D., Ten Eick R. E. Voltage-dependent block of cardiac inward-rectifying potassium current by monovalent cations. J Gen Physiol. 1989 Aug;94(2):349–361. doi: 10.1085/jgp.94.2.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Heginbotham L., Abramson T., MacKinnon R. A functional connection between the pores of distantly related ion channels as revealed by mutant K+ channels. Science. 1992 Nov 13;258(5085):1152–1155. doi: 10.1126/science.1279807. [DOI] [PubMed] [Google Scholar]
  16. Heginbotham L., Lu Z., Abramson T., MacKinnon R. Mutations in the K+ channel signature sequence. Biophys J. 1994 Apr;66(4):1061–1067. doi: 10.1016/S0006-3495(94)80887-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Heginbotham L., MacKinnon R. Conduction properties of the cloned Shaker K+ channel. Biophys J. 1993 Nov;65(5):2089–2096. doi: 10.1016/S0006-3495(93)81244-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hidalgo P., MacKinnon R. Revealing the architecture of a K+ channel pore through mutant cycles with a peptide inhibitor. Science. 1995 Apr 14;268(5208):307–310. doi: 10.1126/science.7716527. [DOI] [PubMed] [Google Scholar]
  19. Hille B., Schwarz W. Potassium channels as multi-ion single-file pores. J Gen Physiol. 1978 Oct;72(4):409–442. doi: 10.1085/jgp.72.4.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ho K., Nichols C. G., Lederer W. J., Lytton J., Vassilev P. M., Kanazirska M. V., Hebert S. C. Cloning and expression of an inwardly rectifying ATP-regulated potassium channel. Nature. 1993 Mar 4;362(6415):31–38. doi: 10.1038/362031a0. [DOI] [PubMed] [Google Scholar]
  21. Kavanaugh M. P., Hurst R. S., Yakel J., Varnum M. D., Adelman J. P., North R. A. Multiple subunits of a voltage-dependent potassium channel contribute to the binding site for tetraethylammonium. Neuron. 1992 Mar;8(3):493–497. doi: 10.1016/0896-6273(92)90277-k. [DOI] [PubMed] [Google Scholar]
  22. Kavanaugh M. P., Varnum M. D., Osborne P. B., Christie M. J., Busch A. E., Adelman J. P., North R. A. Interaction between tetraethylammonium and amino acid residues in the pore of cloned voltage-dependent potassium channels. J Biol Chem. 1991 Apr 25;266(12):7583–7587. [PubMed] [Google Scholar]
  23. Kirsch G. E., Drewe J. A., Taglialatela M., Joho R. H., DeBiasi M., Hartmann H. A., Brown A. M. A single nonpolar residue in the deep pore of related K+ channels acts as a K+:Rb+ conductance switch. Biophys J. 1992 Apr;62(1):136–144. doi: 10.1016/S0006-3495(92)81800-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kirsch G. E., Shieh C. C., Drewe J. A., Vener D. F., Brown A. M. Segmental exchanges define 4-aminopyridine binding and the inner mouth of K+ pores. Neuron. 1993 Sep;11(3):503–512. doi: 10.1016/0896-6273(93)90154-j. [DOI] [PubMed] [Google Scholar]
  25. Kubo Y., Baldwin T. J., Jan Y. N., Jan L. Y. Primary structure and functional expression of a mouse inward rectifier potassium channel. Nature. 1993 Mar 11;362(6416):127–133. doi: 10.1038/362127a0. [DOI] [PubMed] [Google Scholar]
  26. Kubo Y., Reuveny E., Slesinger P. A., Jan Y. N., Jan L. Y. Primary structure and functional expression of a rat G-protein-coupled muscarinic potassium channel. Nature. 1993 Aug 26;364(6440):802–806. doi: 10.1038/364802a0. [DOI] [PubMed] [Google Scholar]
  27. Liman E. R., Tytgat J., Hess P. Subunit stoichiometry of a mammalian K+ channel determined by construction of multimeric cDNAs. Neuron. 1992 Nov;9(5):861–871. doi: 10.1016/0896-6273(92)90239-a. [DOI] [PubMed] [Google Scholar]
  28. Lopatin A. N., Makhina E. N., Nichols C. G. Potassium channel block by cytoplasmic polyamines as the mechanism of intrinsic rectification. Nature. 1994 Nov 24;372(6504):366–369. doi: 10.1038/372366a0. [DOI] [PubMed] [Google Scholar]
  29. Lopez G. A., Jan Y. N., Jan L. Y. Evidence that the S6 segment of the Shaker voltage-gated K+ channel comprises part of the pore. Nature. 1994 Jan 13;367(6459):179–182. doi: 10.1038/367179a0. [DOI] [PubMed] [Google Scholar]
  30. Lu Z., MacKinnon R. A conductance maximum observed in an inward-rectifier potassium channel. J Gen Physiol. 1994 Sep;104(3):477–486. doi: 10.1085/jgp.104.3.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lu Z., MacKinnon R. Electrostatic tuning of Mg2+ affinity in an inward-rectifier K+ channel. Nature. 1994 Sep 15;371(6494):243–246. doi: 10.1038/371243a0. [DOI] [PubMed] [Google Scholar]
  32. Lü Q., Miller C. Silver as a probe of pore-forming residues in a potassium channel. Science. 1995 Apr 14;268(5208):304–307. doi: 10.1126/science.7716526. [DOI] [PubMed] [Google Scholar]
  33. MacKinnon R., Miller C. Mutant potassium channels with altered binding of charybdotoxin, a pore-blocking peptide inhibitor. Science. 1989 Sep 22;245(4924):1382–1385. doi: 10.1126/science.2476850. [DOI] [PubMed] [Google Scholar]
  34. MacKinnon R., Yellen G. Mutations affecting TEA blockade and ion permeation in voltage-activated K+ channels. Science. 1990 Oct 12;250(4978):276–279. doi: 10.1126/science.2218530. [DOI] [PubMed] [Google Scholar]
  35. Matsuda H., Saigusa A., Irisawa H. Ohmic conductance through the inwardly rectifying K channel and blocking by internal Mg2+. Nature. 1987 Jan 8;325(7000):156–159. doi: 10.1038/325156a0. [DOI] [PubMed] [Google Scholar]
  36. Mitra R. L., Morad M. Permeance of Cs+ and Rb+ through the inwardly rectifying K+ channel in guinea pig ventricular myocytes. J Membr Biol. 1991 May;122(1):33–42. doi: 10.1007/BF01872737. [DOI] [PubMed] [Google Scholar]
  37. Ohmori H. Inactivation kinetics and steady-state current noise in the anomalous rectifier of tunicate egg cell membranes. J Physiol. 1978 Aug;281:77–99. doi: 10.1113/jphysiol.1978.sp012410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Pascual J. M., Shieh C. C., Kirsch G. E., Brown A. M. K+ pore structure revealed by reporter cysteines at inner and outer surfaces. Neuron. 1995 May;14(5):1055–1063. doi: 10.1016/0896-6273(95)90344-5. [DOI] [PubMed] [Google Scholar]
  39. Pérez-Cornejo P., Begenisich T. The multi-ion nature of the pore in Shaker K+ channels. Biophys J. 1994 Jun;66(6):1929–1938. doi: 10.1016/S0006-3495(94)80986-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Shieh C. C., Kirsch G. E. Mutational analysis of ion conduction and drug binding sites in the inner mouth of voltage-gated K+ channels. Biophys J. 1994 Dec;67(6):2316–2325. doi: 10.1016/S0006-3495(94)80718-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Shioya T., Matsuda H., Noma A. Fast and slow blockades of the inward-rectifier K+ channel by external divalent cations in guinea-pig cardiac myocytes. Pflugers Arch. 1993 Feb;422(5):427–435. doi: 10.1007/BF00375067. [DOI] [PubMed] [Google Scholar]
  42. Silver M. R., Shapiro M. S., DeCoursey T. E. Effects of external Rb+ on inward rectifier K+ channels of bovine pulmonary artery endothelial cells. J Gen Physiol. 1994 Apr;103(4):519–548. doi: 10.1085/jgp.103.4.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Slesinger P. A., Jan Y. N., Jan L. Y. The S4-S5 loop contributes to the ion-selective pore of potassium channels. Neuron. 1993 Oct;11(4):739–749. doi: 10.1016/0896-6273(93)90083-4. [DOI] [PubMed] [Google Scholar]
  44. Standen N. B., Stanfield P. R. A potential- and time-dependent blockade of inward rectification in frog skeletal muscle fibres by barium and strontium ions. J Physiol. 1978 Jul;280:169–191. doi: 10.1113/jphysiol.1978.sp012379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Standen N. B., Stanfield P. R. Rubidium block and rubidium permeability of the inward rectifier of frog skeletal muscle fibres. J Physiol. 1980 Jul;304:415–435. doi: 10.1113/jphysiol.1980.sp013333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Taglialatela M., Champagne M. S., Drewe J. A., Brown A. M. Comparison of H5, S6, and H5-S6 exchanges on pore properties of voltage-dependent K+ channels. J Biol Chem. 1994 May 13;269(19):13867–13873. [PubMed] [Google Scholar]
  47. Taglialatela M., Drewe J. A., Kirsch G. E., De Biasi M., Hartmann H. A., Brown A. M. Regulation of K+/Rb+ selectivity and internal TEA blockade by mutations at a single site in K+ pores. Pflugers Arch. 1993 Apr;423(1-2):104–112. doi: 10.1007/BF00374967. [DOI] [PubMed] [Google Scholar]
  48. Vandenberg C. A. Inward rectification of a potassium channel in cardiac ventricular cells depends on internal magnesium ions. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2560–2564. doi: 10.1073/pnas.84.8.2560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wagoner P. K., Oxford G. S. Cation permeation through the voltage-dependent potassium channel in the squid axon. Characteristics and mechanisms. J Gen Physiol. 1987 Aug;90(2):261–290. doi: 10.1085/jgp.90.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wang W., Sackin H., Giebisch G. Renal potassium channels and their regulation. Annu Rev Physiol. 1992;54:81–96. doi: 10.1146/annurev.ph.54.030192.000501. [DOI] [PubMed] [Google Scholar]
  51. Woodhull A. M. Ionic blockage of sodium channels in nerve. J Gen Physiol. 1973 Jun;61(6):687–708. doi: 10.1085/jgp.61.6.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Yang J., Jan Y. N., Jan L. Y. Control of rectification and permeation by residues in two distinct domains in an inward rectifier K+ channel. Neuron. 1995 May;14(5):1047–1054. doi: 10.1016/0896-6273(95)90343-7. [DOI] [PubMed] [Google Scholar]
  53. Yellen G., Jurman M. E., Abramson T., MacKinnon R. Mutations affecting internal TEA blockade identify the probable pore-forming region of a K+ channel. Science. 1991 Feb 22;251(4996):939–942. doi: 10.1126/science.2000494. [DOI] [PubMed] [Google Scholar]
  54. Yool A. J., Schwarz T. L. Alteration of ionic selectivity of a K+ channel by mutation of the H5 region. Nature. 1991 Feb 21;349(6311):700–704. doi: 10.1038/349700a0. [DOI] [PubMed] [Google Scholar]

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