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. 1996 Dec 15;98(12):2874–2886. doi: 10.1172/JCI119116

Local anesthetics as effectors of allosteric gating. Lidocaine effects on inactivation-deficient rat skeletal muscle Na channels.

J R Balser 1, H B Nuss 1, D W Orias 1, D C Johns 1, E Marban 1, G F Tomaselli 1, J H Lawrence 1
PMCID: PMC507755  PMID: 8981936

Abstract

Time- and voltage-dependent local anesthetic effects on sodium (Na) currents are generally interpreted using modulated receptor models that require formation of drug-associated nonconducting states with high affinity for the inactivated channel. The availability of inactivation-deficient Na channels has enabled us to test this traditional view of the drug-channel interaction. Rat skeletal muscle Na channels were mutated in the III-IV linker to disable fast inactivation (F1304Q: FQ). Lidocaine accelerated the decay of whole-cell FQ currents in Xenopus oocytes, reestablishing the wild-type phenotype; peak inward current at -20 mV was blocked with an IC50 of 513 microM, while plateau current was blocked with an IC50 of only 74 microM (P < 0.005 vs. peak). In single-channel experiments, mean open time was unaltered and unitary current was only reduced at higher drug concentrations, suggesting that open-channel block does not explain the effect of lidocaine on FQ plateau current. We considered a simple model in which lidocaine reduced the free energy for inactivation, causing altered coupling between activation and inactivation. This model readily simulated macroscopic Na current kinetics over a range of lidocaine concentrations. Traditional modulated receptor models which did not modify coupling between gating processes could not reproduce the effects of lidocaine with rate constants constrained by single-channel data. Our results support a reinterpretation of local anesthetic action whereby lidocaine functions as an allosteric effector to enhance Na channel inactivation.

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Selected References

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  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. Armstrong C. M. Interaction of tetraethylammonium ion derivatives with the potassium channels of giant axons. J Gen Physiol. 1971 Oct;58(4):413–437. doi: 10.1085/jgp.58.4.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Backx P. H., Yue D. T., Lawrence J. H., Marban E., Tomaselli G. F. Molecular localization of an ion-binding site within the pore of mammalian sodium channels. Science. 1992 Jul 10;257(5067):248–251. doi: 10.1126/science.1321496. [DOI] [PubMed] [Google Scholar]
  4. Balser J. R., Nuss H. B., Romashko D. N., Marban E., Tomaselli G. F. Functional consequences of lidocaine binding to slow-inactivated sodium channels. J Gen Physiol. 1996 May;107(5):643–658. doi: 10.1085/jgp.107.5.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Balser J. R., Roden D. M., Bennett P. B. Global parameter optimization for cardiac potassium channel gating models. Biophys J. 1990 Mar;57(3):433–444. doi: 10.1016/S0006-3495(90)82560-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bean B. P., Cohen C. J., Tsien R. W. Lidocaine block of cardiac sodium channels. J Gen Physiol. 1983 May;81(5):613–642. doi: 10.1085/jgp.81.5.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bennett P. B., Jr, Makita N., George A. L., Jr A molecular basis for gating mode transitions in human skeletal muscle Na+ channels. FEBS Lett. 1993 Jul 12;326(1-3):21–24. doi: 10.1016/0014-5793(93)81752-l. [DOI] [PubMed] [Google Scholar]
  8. Bennett P. B., Valenzuela C., Chen L. Q., Kallen R. G. On the molecular nature of the lidocaine receptor of cardiac Na+ channels. Modification of block by alterations in the alpha-subunit III-IV interdomain. Circ Res. 1995 Sep;77(3):584–592. doi: 10.1161/01.res.77.3.584. [DOI] [PubMed] [Google Scholar]
  9. Bennett P. B., Yazawa K., Makita N., George A. L., Jr Molecular mechanism for an inherited cardiac arrhythmia. Nature. 1995 Aug 24;376(6542):683–685. doi: 10.1038/376683a0. [DOI] [PubMed] [Google Scholar]
  10. Cahalan M. D., Almers W. Interactions between quaternary lidocaine, the sodium channel gates, and tetrodotoxin. Biophys J. 1979 Jul;27(1):39–55. doi: 10.1016/S0006-3495(79)85201-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cahalan M. D. Local anesthetic block of sodium channels in normal and pronase-treated squid giant axons. Biophys J. 1978 Aug;23(2):285–311. doi: 10.1016/S0006-3495(78)85449-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Chahine M., George A. L., Jr, Zhou M., Ji S., Sun W., Barchi R. L., Horn R. Sodium channel mutations in paramyotonia congenita uncouple inactivation from activation. Neuron. 1994 Feb;12(2):281–294. doi: 10.1016/0896-6273(94)90271-2. [DOI] [PubMed] [Google Scholar]
  14. Chang S. Y., Satin J., Fozzard H. A. Modal behavior of the mu 1 Na+ channel and effects of coexpression of the beta 1-subunit. Biophys J. 1996 Jun;70(6):2581–2592. doi: 10.1016/S0006-3495(96)79829-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Courtney K. R. Mechanism of frequency-dependent inhibition of sodium currents in frog myelinated nerve by the lidocaine derivative GEA. J Pharmacol Exp Ther. 1975 Nov;195(2):225–236. [PubMed] [Google Scholar]
  16. Dumaine R., Wang Q., Keating M. T., Hartmann H. A., Schwartz P. J., Brown A. M., Kirsch G. E. Multiple mechanisms of Na+ channel--linked long-QT syndrome. Circ Res. 1996 May;78(5):916–924. doi: 10.1161/01.res.78.5.916. [DOI] [PubMed] [Google Scholar]
  17. Gingrich K. J., Beardsley D., Yue D. T. Ultra-deep blockade of Na+ channels by a quaternary ammonium ion: catalysis by a transition-intermediate state? J Physiol. 1993 Nov;471:319–341. doi: 10.1113/jphysiol.1993.sp019903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Grant A. O., Dietz M. A., Gilliam F. R., 3rd, Starmer C. F. Blockade of cardiac sodium channels by lidocaine. Single-channel analysis. Circ Res. 1989 Nov;65(5):1247–1262. doi: 10.1161/01.res.65.5.1247. [DOI] [PubMed] [Google Scholar]
  19. HODGKIN A. L., HUXLEY A. F. A quantitative description of membrane current and its application to conduction and excitation in nerve. J Physiol. 1952 Aug;117(4):500–544. doi: 10.1113/jphysiol.1952.sp004764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Hille B. Local anesthetics: hydrophilic and hydrophobic pathways for the drug-receptor reaction. J Gen Physiol. 1977 Apr;69(4):497–515. doi: 10.1085/jgp.69.4.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hondeghem L. M., Katzung B. G. Time- and voltage-dependent interactions of antiarrhythmic drugs with cardiac sodium channels. Biochim Biophys Acta. 1977 Nov 14;472(3-4):373–398. doi: 10.1016/0304-4157(77)90003-x. [DOI] [PubMed] [Google Scholar]
  24. Horn R., Patlak J., Stevens C. F. Sodium channels need not open before they inactivate. Nature. 1981 Jun 4;291(5814):426–427. doi: 10.1038/291426a0. [DOI] [PubMed] [Google Scholar]
  25. Horn R., Vandenberg C. A. Statistical properties of single sodium channels. J Gen Physiol. 1984 Oct;84(4):505–534. doi: 10.1085/jgp.84.4.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  31. Kuo C. C., Bean B. P. Na+ channels must deactivate to recover from inactivation. Neuron. 1994 Apr;12(4):819–829. doi: 10.1016/0896-6273(94)90335-2. [DOI] [PubMed] [Google Scholar]
  32. Lawrence J. H., Orias D. W., Balser J. R., Nuss H. B., Tomaselli G. F., O'Rourke B., Marban E. Single-channel analysis of inactivation-defective rat skeletal muscle sodium channels containing the F1304Q mutation. Biophys J. 1996 Sep;71(3):1285–1294. doi: 10.1016/S0006-3495(96)79329-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lawrence J. H., Yue D. T., Rose W. C., Marban E. Sodium channel inactivation from resting states in guinea-pig ventricular myocytes. J Physiol. 1991 Nov;443:629–650. doi: 10.1113/jphysiol.1991.sp018855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. MONOD J., WYMAN J., CHANGEUX J. P. ON THE NATURE OF ALLOSTERIC TRANSITIONS: A PLAUSIBLE MODEL. J Mol Biol. 1965 May;12:88–118. doi: 10.1016/s0022-2836(65)80285-6. [DOI] [PubMed] [Google Scholar]
  35. Makielski J. C., Limberis J. T., Chang S. Y., Fan Z., Kyle J. W. Coexpression of beta 1 with cardiac sodium channel alpha subunits in oocytes decreases lidocaine block. Mol Pharmacol. 1996 Jan;49(1):30–39. [PubMed] [Google Scholar]
  36. Marks T. N., Jones S. W. Calcium currents in the A7r5 smooth muscle-derived cell line. An allosteric model for calcium channel activation and dihydropyridine agonist action. J Gen Physiol. 1992 Mar;99(3):367–390. doi: 10.1085/jgp.99.3.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Matsubara T., Clarkson C., Hondeghem L. Lidocaine blocks open and inactivated cardiac sodium channels. Naunyn Schmiedebergs Arch Pharmacol. 1987 Aug;336(2):224–231. doi: 10.1007/BF00165809. [DOI] [PubMed] [Google Scholar]
  38. McPhee J. C., Ragsdale D. S., Scheuer T., Catterall W. A. A critical role for transmembrane segment IVS6 of the sodium channel alpha subunit in fast inactivation. J Biol Chem. 1995 May 19;270(20):12025–12034. doi: 10.1074/jbc.270.20.12025. [DOI] [PubMed] [Google Scholar]
  39. Moorman J. R., Kirsch G. E., Brown A. M., Joho R. H. Changes in sodium channel gating produced by point mutations in a cytoplasmic linker. Science. 1990 Nov 2;250(4981):688–691. doi: 10.1126/science.2173138. [DOI] [PubMed] [Google Scholar]
  40. Moorman J. R., Kirsch G. E., VanDongen A. M., Joho R. H., Brown A. M. Fast and slow gating of sodium channels encoded by a single mRNA. Neuron. 1990 Feb;4(2):243–252. doi: 10.1016/0896-6273(90)90099-2. [DOI] [PubMed] [Google Scholar]
  41. Neher E., Steinbach J. H. Local anaesthetics transiently block currents through single acetylcholine-receptor channels. J Physiol. 1978 Apr;277:153–176. doi: 10.1113/jphysiol.1978.sp012267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Nilius B., Benndorf K., Markwardt F. Effects of lidocaine on single cardiac sodium channels. J Mol Cell Cardiol. 1987 Sep;19(9):865–874. doi: 10.1016/s0022-2828(87)80615-6. [DOI] [PubMed] [Google Scholar]
  43. Nuss H. B., Balser J. R., Orias D. W., Lawrence J. H., Tomaselli G. F., Marban E. Coupling between fast and slow inactivation revealed by analysis of a point mutation (F1304Q) in mu 1 rat skeletal muscle sodium channels. J Physiol. 1996 Jul 15;494(Pt 2):411–429. doi: 10.1113/jphysiol.1996.sp021502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Nuss H. B., Chiamvimonvat N., Pérez-García M. T., Tomaselli G. F., Marbán E. Functional association of the beta 1 subunit with human cardiac (hH1) and rat skeletal muscle (mu 1) sodium channel alpha subunits expressed in Xenopus oocytes. J Gen Physiol. 1995 Dec;106(6):1171–1191. doi: 10.1085/jgp.106.6.1171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Nuss H. B., Tomaselli G. F., Marbán E. Cardiac sodium channels (hH1) are intrinsically more sensitive to block by lidocaine than are skeletal muscle (mu 1) channels. J Gen Physiol. 1995 Dec;106(6):1193–1209. doi: 10.1085/jgp.106.6.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. O'Leary M. E., Kallen R. G., Horn R. Evidence for a direct interaction between internal tetra-alkylammonium cations and the inactivation gate of cardiac sodium channels. J Gen Physiol. 1994 Sep;104(3):523–539. doi: 10.1085/jgp.104.3.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Patlak J. Molecular kinetics of voltage-dependent Na+ channels. Physiol Rev. 1991 Oct;71(4):1047–1080. doi: 10.1152/physrev.1991.71.4.1047. [DOI] [PubMed] [Google Scholar]
  48. 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]
  49. Ragsdale D. S., McPhee J. C., Scheuer T., Catterall W. A. Molecular determinants of state-dependent block of Na+ channels by local anesthetics. Science. 1994 Sep 16;265(5179):1724–1728. doi: 10.1126/science.8085162. [DOI] [PubMed] [Google Scholar]
  50. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Schwarz W., Palade P. T., Hille B. Local anesthetics. Effect of pH on use-dependent block of sodium channels in frog muscle. Biophys J. 1977 Dec;20(3):343–368. doi: 10.1016/S0006-3495(77)85554-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Strichartz G. R. The inhibition of sodium currents in myelinated nerve by quaternary derivatives of lidocaine. J Gen Physiol. 1973 Jul;62(1):37–57. doi: 10.1085/jgp.62.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Stühmer W., Conti F., Suzuki H., Wang X. D., Noda M., Yahagi N., Kubo H., Numa S. Structural parts involved in activation and inactivation of the sodium channel. Nature. 1989 Jun 22;339(6226):597–603. doi: 10.1038/339597a0. [DOI] [PubMed] [Google Scholar]
  54. Tomaselli G. F., Chiamvimonvat N., Nuss H. B., Balser J. R., Pérez-García M. T., Xu R. H., Orias D. W., Backx P. H., Marban E. A mutation in the pore of the sodium channel alters gating. Biophys J. 1995 May;68(5):1814–1827. doi: 10.1016/S0006-3495(95)80358-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. 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]
  56. Vassilev P. M., Scheuer T., Catterall W. A. Identification of an intracellular peptide segment involved in sodium channel inactivation. Science. 1988 Sep 23;241(4873):1658–1661. doi: 10.1126/science.241.4873.1658. [DOI] [PubMed] [Google Scholar]
  57. Vassilev P., Scheuer T., Catterall W. A. Inhibition of inactivation of single sodium channels by a site-directed antibody. Proc Natl Acad Sci U S A. 1989 Oct;86(20):8147–8151. doi: 10.1073/pnas.86.20.8147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. 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]
  59. 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]
  60. Yeh J. Z. Sodium inactivation mechanism modulates QX-314 block of sodium channels in squid axons. Biophys J. 1978 Nov;24(2):569–574. doi: 10.1016/S0006-3495(78)85403-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Zamponi G. W., Doyle D. D., French R. J. Fast lidocaine block of cardiac and skeletal muscle sodium channels: one site with two routes of access. Biophys J. 1993 Jul;65(1):80–90. doi: 10.1016/S0006-3495(93)81042-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Zhou J. Y., Potts J. F., Trimmer J. S., Agnew W. S., Sigworth F. J. Multiple gating modes and the effect of modulating factors on the microI sodium channel. Neuron. 1991 Nov;7(5):775–785. doi: 10.1016/0896-6273(91)90280-d. [DOI] [PubMed] [Google Scholar]
  63. de Leon M., Wang Y., Jones L., Perez-Reyes E., Wei X., Soong T. W., Snutch T. P., Yue D. T. Essential Ca(2+)-binding motif for Ca(2+)-sensitive inactivation of L-type Ca2+ channels. Science. 1995 Dec 1;270(5241):1502–1506. doi: 10.1126/science.270.5241.1502. [DOI] [PubMed] [Google Scholar]

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