Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 2001 May;80(5):2221–2230. doi: 10.1016/S0006-3495(01)76195-4

A single residue differentiates between human cardiac and skeletal muscle Na+ channel slow inactivation.

Y Y Vilin 1, E Fujimoto 1, P C Ruben 1
PMCID: PMC1301414  PMID: 11325725

Abstract

Slow inactivation determines the availability of voltage-gated sodium channels during prolonged depolarization. Slow inactivation in hNa(V)1.4 channels occurs with a higher probability than hNa(V)1.5 sodium channels; however, the precise molecular mechanism for this difference remains unclear. Using the macropatch technique we show that the DII S5-S6 p-region uniquely confers the probability of slow inactivation from parental hNa(V)1.5 and hNa(V)1.4 channels into chimerical constructs expressed in Xenopus oocytes. Site-directed mutagenesis was used to test whether a specific region within DII S5-S6 controls the probability of slow inactivation. We found that substituting V754 in hNa(V)1.4 with isoleucine from the corresponding position (891) in hNa(V)1.5 produced steady-state slow inactivation statistically indistinguishable from that in wild-type hNa(V)1.5 channels, whereas other mutations have little or no effect on slow inactivation. This result indicates that residues V754 in hNa(V)1.4 and I891in hNa(V)1.5 are unique in determining the probability of slow inactivation characteristic of these isoforms. Exchanging S5-S6 linkers between hNa(V)1.4 and hNa(V)1.5 channels had no consistent effect on the voltage-dependent slow time inactivation constants [tau(V)]. This suggests that the molecular structures regulating rates of entry into and exit from the slow inactivated state are different from those controlling the steady-state probability and reside outside the p-regions.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Armstrong C. M., Bezanilla F. Inactivation of the sodium channel. II. Gating current experiments. J Gen Physiol. 1977 Nov;70(5):567–590. doi: 10.1085/jgp.70.5.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bezanilla F., Taylor R. E., Fernández J. M. Distribution and kinetics of membrane dielectric polarization. 1. Long-term inactivation of gating currents. J Gen Physiol. 1982 Jan;79(1):21–40. doi: 10.1085/jgp.79.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boland L. M., Jurman M. E., Yellen G. Cysteines in the Shaker K+ channel are not essential for channel activity or zinc modulation. Biophys J. 1994 Mar;66(3 Pt 1):694–699. doi: 10.1016/s0006-3495(94)80843-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cannon S. C., McClatchey A. I., Gusella J. F. Modification of the Na+ current conducted by the rat skeletal muscle alpha subunit by coexpression with a human brain beta subunit. Pflugers Arch. 1993 Apr;423(1-2):155–157. doi: 10.1007/BF00374974. [DOI] [PubMed] [Google Scholar]
  5. Catterall W. A. Structure and function of voltage-gated ion channels. Trends Neurosci. 1993 Dec;16(12):500–506. doi: 10.1016/0166-2236(93)90193-p. [DOI] [PubMed] [Google Scholar]
  6. Cha A., Bezanilla F. Characterizing voltage-dependent conformational changes in the Shaker K+ channel with fluorescence. Neuron. 1997 Nov;19(5):1127–1140. doi: 10.1016/s0896-6273(00)80403-1. [DOI] [PubMed] [Google Scholar]
  7. Cha A., Ruben P. C., George A. L., Jr, Fujimoto E., Bezanilla F. Voltage sensors in domains III and IV, but not I and II, are immobilized by Na+ channel fast inactivation. Neuron. 1999 Jan;22(1):73–87. doi: 10.1016/s0896-6273(00)80680-7. [DOI] [PubMed] [Google Scholar]
  8. Chandler W. K., Meves H. Slow changes in membrane permeability and long-lasting action potentials in axons perfused with fluoride solutions. J Physiol. 1970 Dec;211(3):707–728. doi: 10.1113/jphysiol.1970.sp009300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chen C., Cannon S. C. Modulation of Na+ channel inactivation by the beta 1 subunit: a deletion analysis. Pflugers Arch. 1995 Dec;431(2):186–195. doi: 10.1007/BF00410190. [DOI] [PubMed] [Google Scholar]
  10. Cummins T. R., Sigworth F. J. Impaired slow inactivation in mutant sodium channels. Biophys J. 1996 Jul;71(1):227–236. doi: 10.1016/S0006-3495(96)79219-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Featherstone D. E., Richmond J. E., Ruben P. C. Interaction between fast and slow inactivation in Skm1 sodium channels. Biophys J. 1996 Dec;71(6):3098–3109. doi: 10.1016/S0006-3495(96)79504-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fleidervish I. A., Friedman A., Gutnick M. J. Slow inactivation of Na+ current and slow cumulative spike adaptation in mouse and guinea-pig neocortical neurones in slices. J Physiol. 1996 May 15;493(Pt 1):83–97. doi: 10.1113/jphysiol.1996.sp021366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fozzard H. A., Hanck D. A. Structure and function of voltage-dependent sodium channels: comparison of brain II and cardiac isoforms. Physiol Rev. 1996 Jul;76(3):887–926. doi: 10.1152/physrev.1996.76.3.887. [DOI] [PubMed] [Google Scholar]
  14. Gellens M. E., George A. L., Jr, Chen L. Q., Chahine M., Horn R., Barchi R. L., Kallen R. G. Primary structure and functional expression of the human cardiac tetrodotoxin-insensitive voltage-dependent sodium channel. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):554–558. doi: 10.1073/pnas.89.2.554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. George A. L., Jr, Komisarof J., Kallen R. G., Barchi R. L. Primary structure of the adult human skeletal muscle voltage-dependent sodium channel. Ann Neurol. 1992 Feb;31(2):131–137. doi: 10.1002/ana.410310203. [DOI] [PubMed] [Google Scholar]
  16. Goldin A. L., Barchi R. L., Caldwell J. H., Hofmann F., Howe J. R., Hunter J. C., Kallen R. G., Mandel G., Meisler M. H., Netter Y. B. Nomenclature of voltage-gated sodium channels. Neuron. 2000 Nov;28(2):365–368. doi: 10.1016/s0896-6273(00)00116-1. [DOI] [PubMed] [Google Scholar]
  17. Groome J. R., Fujimoto E., George A. L., Ruben P. C. Differential effects of homologous S4 mutations in human skeletal muscle sodium channels on deactivation gating from open and inactivated states. J Physiol. 1999 May 1;516(Pt 3):687–698. doi: 10.1111/j.1469-7793.1999.0687u.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Groome J. R., Fujimoto E., Ruben P. C. The delay in recovery from fast inactivation in skeletal muscle sodium channels is deactivation. Cell Mol Neurobiol. 2000 Aug;20(4):521–527. doi: 10.1023/A:1007040731407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hartmann H. A., Tiedeman A. A., Chen S. F., Brown A. M., Kirsch G. E. Effects of III-IV linker mutations on human heart Na+ channel inactivation gating. Circ Res. 1994 Jul;75(1):114–122. doi: 10.1161/01.res.75.1.114. [DOI] [PubMed] [Google Scholar]
  20. Hayward L. J., Brown R. H., Jr, Cannon S. C. Slow inactivation differs among mutant Na channels associated with myotonia and periodic paralysis. Biophys J. 1997 Mar;72(3):1204–1219. doi: 10.1016/S0006-3495(97)78768-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [PubMed] [Google Scholar]
  22. Hoshi T., Zagotta W. N., Aldrich R. W. Biophysical and molecular mechanisms of Shaker potassium channel inactivation. Science. 1990 Oct 26;250(4980):533–538. doi: 10.1126/science.2122519. [DOI] [PubMed] [Google Scholar]
  23. Hoshi T., Zagotta W. N., Aldrich R. W. Two types of inactivation in Shaker K+ channels: effects of alterations in the carboxy-terminal region. Neuron. 1991 Oct;7(4):547–556. doi: 10.1016/0896-6273(91)90367-9. [DOI] [PubMed] [Google Scholar]
  24. Isom L. L., De Jongh K. S., Catterall W. A. Auxiliary subunits of voltage-gated ion channels. Neuron. 1994 Jun;12(6):1183–1194. doi: 10.1016/0896-6273(94)90436-7. [DOI] [PubMed] [Google Scholar]
  25. Isom L. L., De Jongh K. S., Patton D. E., Reber B. F., Offord J., Charbonneau H., Walsh K., Goldin A. L., Catterall W. A. Primary structure and functional expression of the beta 1 subunit of the rat brain sodium channel. Science. 1992 May 8;256(5058):839–842. doi: 10.1126/science.1375395. [DOI] [PubMed] [Google Scholar]
  26. Ji S., George A. L., Jr, Horn R., Barchi R. L. Paramyotonia congenita mutations reveal different roles for segments S3 and S4 of domain D4 in hSkM1 sodium channel gating. J Gen Physiol. 1996 Feb;107(2):183–194. doi: 10.1085/jgp.107.2.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kiss L., LoTurco J., Korn S. J. Contribution of the selectivity filter to inactivation in potassium channels. Biophys J. 1999 Jan;76(1 Pt 1):253–263. doi: 10.1016/S0006-3495(99)77194-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kontis K. J., Goldin A. L. Sodium channel inactivation is altered by substitution of voltage sensor positive charges. J Gen Physiol. 1997 Oct;110(4):403–413. doi: 10.1085/jgp.110.4.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Liu Y., Jurman M. E., Yellen G. Dynamic rearrangement of the outer mouth of a K+ channel during gating. Neuron. 1996 Apr;16(4):859–867. doi: 10.1016/s0896-6273(00)80106-3. [DOI] [PubMed] [Google Scholar]
  30. Loots E., Isacoff E. Y. Molecular coupling of S4 to a K(+) channel's slow inactivation gate. J Gen Physiol. 2000 Nov;116(5):623–636. doi: 10.1085/jgp.116.5.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Loots E., Isacoff E. Y. Protein rearrangements underlying slow inactivation of the Shaker K+ channel. J Gen Physiol. 1998 Oct;112(4):377–389. doi: 10.1085/jgp.112.4.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Makita N., Bennett P. B., George A. L., Jr Molecular determinants of beta 1 subunit-induced gating modulation in voltage-dependent Na+ channels. J Neurosci. 1996 Nov 15;16(22):7117–7127. doi: 10.1523/JNEUROSCI.16-22-07117.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Makita N., Bennett P. B., Jr, George A. L., Jr Voltage-gated Na+ channel beta 1 subunit mRNA expressed in adult human skeletal muscle, heart, and brain is encoded by a single gene. J Biol Chem. 1994 Mar 11;269(10):7571–7578. [PubMed] [Google Scholar]
  34. Marban E., Yamagishi T., Tomaselli G. F. Structure and function of voltage-gated sodium channels. J Physiol. 1998 May 1;508(Pt 3):647–657. doi: 10.1111/j.1469-7793.1998.647bp.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McCormick K. A., Isom L. L., Ragsdale D., Smith D., Scheuer T., Catterall W. A. Molecular determinants of Na+ channel function in the extracellular domain of the beta1 subunit. J Biol Chem. 1998 Feb 13;273(7):3954–3962. doi: 10.1074/jbc.273.7.3954. [DOI] [PubMed] [Google Scholar]
  36. Mitrovic N., George A. L., Jr, Horn R. Role of domain 4 in sodium channel slow inactivation. J Gen Physiol. 2000 Jun;115(6):707–718. doi: 10.1085/jgp.115.6.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. O'Reilly J. P., Wang S. Y., Kallen R. G., Wang G. K. Comparison of slow inactivation in human heart and rat skeletal muscle Na+ channel chimaeras. J Physiol. 1999 Feb 15;515(Pt 1):61–73. doi: 10.1111/j.1469-7793.1999.061ad.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Patton D. E., Isom L. L., Catterall W. A., Goldin A. L. The adult rat brain beta 1 subunit modifies activation and inactivation gating of multiple sodium channel alpha subunits. J Biol Chem. 1994 Jul 1;269(26):17649–17655. [PubMed] [Google Scholar]
  39. Patton D. E., West J. W., Catterall W. A., Goldin A. L. Amino acid residues required for fast Na(+)-channel inactivation: charge neutralizations and deletions in the III-IV linker. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10905–10909. doi: 10.1073/pnas.89.22.10905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Richmond J. E., Featherstone D. E., Hartmann H. A., Ruben P. C. Slow inactivation in human cardiac sodium channels. Biophys J. 1998 Jun;74(6):2945–2952. doi: 10.1016/S0006-3495(98)78001-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rogart R. B., Cribbs L. L., Muglia L. K., Kephart D. D., Kaiser M. W. Molecular cloning of a putative tetrodotoxin-resistant rat heart Na+ channel isoform. Proc Natl Acad Sci U S A. 1989 Oct;86(20):8170–8174. doi: 10.1073/pnas.86.20.8170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ruben P. C., Starkus J. G., Rayner M. D. Holding potential affects the apparent voltage-sensitivity of sodium channel activation in crayfish giant axons. Biophys J. 1990 Nov;58(5):1169–1181. doi: 10.1016/S0006-3495(90)82458-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ruben P. C., Starkus J. G., Rayner M. D. Steady-state availability of sodium channels. Interactions between activation and slow inactivation. Biophys J. 1992 Apr;61(4):941–955. doi: 10.1016/S0006-3495(92)81901-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rudy B. Slow inactivation of the sodium conductance in squid giant axons. Pronase resistance. J Physiol. 1978 Oct;283:1–21. doi: 10.1113/jphysiol.1978.sp012485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Ruff R. L., Simoncini L., Stühmer W. Slow sodium channel inactivation in mammalian muscle: a possible role in regulating excitability. Muscle Nerve. 1988 May;11(5):502–510. doi: 10.1002/mus.880110514. [DOI] [PubMed] [Google Scholar]
  46. Ruff R. L. Single-channel basis of slow inactivation of Na+ channels in rat skeletal muscle. Am J Physiol. 1996 Sep;271(3 Pt 1):C971–C981. doi: 10.1152/ajpcell.1996.271.3.C971. [DOI] [PubMed] [Google Scholar]
  47. Salgado V. L., Yeh J. Z., Narahashi T. Voltage-dependent removal of sodium inactivation by N-bromoacetamide and pronase. Biophys J. 1985 Apr;47(4):567–571. doi: 10.1016/S0006-3495(85)83952-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Sato C., Sato M., Iwasaki A., Doi T., Engel A. The sodium channel has four domains surrounding a central pore. J Struct Biol. 1998;121(3):314–325. doi: 10.1006/jsbi.1998.3990. [DOI] [PubMed] [Google Scholar]
  49. Sawczuk A., Powers R. K., Binder M. D. Spike frequency adaptation studied in hypoglossal motoneurons of the rat. J Neurophysiol. 1995 May;73(5):1799–1810. doi: 10.1152/jn.1995.73.5.1799. [DOI] [PubMed] [Google Scholar]
  50. Sheets M. F., Kyle J. W., Hanck D. A. The role of the putative inactivation lid in sodium channel gating current immobilization. J Gen Physiol. 2000 May;115(5):609–620. doi: 10.1085/jgp.115.5.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Starkus J. G., Shrager P. Modification of slow sodium inactivation in nerve after internal perfusion with trypsin. Am J Physiol. 1978 Nov;235(5):C238–C244. doi: 10.1152/ajpcell.1978.235.5.C238. [DOI] [PubMed] [Google Scholar]
  52. Toib A., Lyakhov V., Marom S. Interaction between duration of activity and time course of recovery from slow inactivation in mammalian brain Na+ channels. J Neurosci. 1998 Mar 1;18(5):1893–1903. doi: 10.1523/JNEUROSCI.18-05-01893.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Townsend C., Horn R. Effect of alkali metal cations on slow inactivation of cardiac Na+ channels. J Gen Physiol. 1997 Jul;110(1):23–33. doi: 10.1085/jgp.110.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Trimmer J. S., Cooperman S. S., Tomiko S. A., Zhou J. Y., Crean S. M., Boyle M. B., Kallen R. G., Sheng Z. H., Barchi R. L., Sigworth F. J. Primary structure and functional expression of a mammalian skeletal muscle sodium channel. Neuron. 1989 Jul;3(1):33–49. doi: 10.1016/0896-6273(89)90113-x. [DOI] [PubMed] [Google Scholar]
  55. Valenzuela C., Bennett P. B., Jr Gating of cardiac Na+ channels in excised membrane patches after modification by alpha-chymotrypsin. Biophys J. 1994 Jul;67(1):161–171. doi: 10.1016/S0006-3495(94)80465-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Vedantham V., Cannon S. C. Slow inactivation does not affect movement of the fast inactivation gate in voltage-gated Na+ channels. J Gen Physiol. 1998 Jan;111(1):83–93. doi: 10.1085/jgp.111.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Vilin Y. Y., Makita N., George A. L., Jr, Ruben P. C. Structural determinants of slow inactivation in human cardiac and skeletal muscle sodium channels. Biophys J. 1999 Sep;77(3):1384–1393. doi: 10.1016/S0006-3495(99)76987-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wang S. Y., Wang G. K. A mutation in segment I-S6 alters slow inactivation of sodium channels. Biophys J. 1997 Apr;72(4):1633–1640. doi: 10.1016/S0006-3495(97)78809-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. West J. W., Patton D. E., Scheuer T., Wang Y., Goldin A. L., Catterall W. A. A cluster of hydrophobic amino acid residues required for fast Na(+)-channel inactivation. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10910–10914. doi: 10.1073/pnas.89.22.10910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Yang J. S., Bennett P. B., Makita N., George A. L., Barchi R. L. Expression of the sodium channel beta 1 subunit in rat skeletal muscle is selectively associated with the tetrodotoxin-sensitive alpha subunit isoform. Neuron. 1993 Nov;11(5):915–922. doi: 10.1016/0896-6273(93)90121-7. [DOI] [PubMed] [Google Scholar]
  61. Yang N., George A. L., Jr, Horn R. Probing the outer vestibule of a sodium channel voltage sensor. Biophys J. 1997 Nov;73(5):2260–2268. doi: 10.1016/S0006-3495(97)78258-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Yang Y., Yan Y., Sigworth F. J. How does the W434F mutation block current in Shaker potassium channels? J Gen Physiol. 1997 Jun;109(6):779–789. doi: 10.1085/jgp.109.6.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Yellen G., Sodickson D., Chen T. Y., Jurman M. E. An engineered cysteine in the external mouth of a K+ channel allows inactivation to be modulated by metal binding. Biophys J. 1994 Apr;66(4):1068–1075. doi: 10.1016/S0006-3495(94)80888-4. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES