title: Modulation of a voltage-gated Na+ channel by sevoflurane involves multiple sites and distinct mechanisms.
abstract: Halogenated inhaled general anesthetic agents modulate voltage-gated ion channels, but the underlying molecular mechanisms are not understood. Many general anesthetic agents regulate voltage-gated Na(+) (NaV) channels, including the commonly used drug sevoflurane. Here, we investigated the putative binding sites and molecular mechanisms of sevoflurane action on the bacterial NaV channel NaChBac by using a combination of molecular dynamics simulation, electrophysiology, and kinetic analysis. Structural modeling revealed multiple sevoflurane interaction sites possibly associated with NaChBac modulation. Electrophysiologically, sevoflurane favors activation and inactivation at low concentrations (0.2 mM), and additionally accelerates current decay at high concentrations (2 mM). Explaining these observations, kinetic modeling suggests concurrent destabilization of closed states and low-affinity open channel block. We propose that the multiple effects of sevoflurane on NaChBac result from simultaneous interactions at multiple sites with distinct affinities. This multiple-site, multiple-mode hypothesis offers a framework to study the structural basis of general anesthetic action.
authors.affiliation: Department of Neuroscience, Thomas Jefferson University, Philadelphia, PA 19107.
authors.forename: Annika F
authors.initials: AF
authors.lastname: Barber
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Carnevale
authors.forename: Michael L
authors.initials: ML
authors.lastname: Klein
authors.forename: Roderic G
authors.initials: RG
authors.lastname: Eckenhoff
authors.forename: Manuel
authors.initials: M
authors.lastname: Covarrubias
mesh_terms: D018685:Anesthetics, Inhalation / Q000378:metabolism / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*; D001665:Binding Sites; D055724:Electrophysiological Phenomena; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects; D007700:Kinetics; D008738:Methyl Ethers / Q000378:metabolism / Q000494:pharmacology*; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D018408:Patch-Clamp Techniques; D011487:Protein Conformation / Q000187:drug effects; D011994:Recombinant Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D000077149:Sevoflurane; D015222:Sodium Channels / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D018685:Anesthetics, Inhalation; D001426:Bacterial Proteins; D008738:Methyl Ethers; C443666:NaChBac protein, bacteria; D011994:Recombinant Proteins; D015222:Sodium Channels; D000077149:Sevoflurane
keywords: MD simulations; anesthesia; anesthetics; membrane proteins
references.citation: Proc Natl Acad Sci U S A. 2010 Aug 10;107(32):14122-7
references.pmid: 20660787
references.citation: Nature. 2012 May 20;486(7401):130-4
references.pmid: 22678295
references.citation: Br J Anaesth. 2013 Apr;110(4):592-9
references.pmid: 23213036
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):15142-7
references.pmid: 18809926
references.citation: Nature. 2011 Jul 10;475(7356):353-8
references.pmid: 21743477
references.citation: J Biol Chem. 2004 Mar 5;279(10):9532-8
references.pmid: 14665618
references.citation: J Biol Chem. 2012 Nov 23;287(48):40425-32
references.pmid: 23038249
references.citation: Nat Neurosci. 1999 May;2(5):422-6
references.pmid: 10321245
references.citation: Proc Natl Acad Sci U S A. 2004 Dec 21;101(51):17873-8
references.pmid: 15583130
references.citation: Anesthesiology. 1998 Apr;88(4):1043-54
references.pmid: 9579514
references.citation: J Biol Chem. 2010 Feb 5;285(6):3685-3694
references.pmid: 19959480
references.citation: Nat Commun. 2013;4:1697
references.pmid: 23591864
references.citation: J Neurochem. 2010 Jun;113(6):1611-20
references.pmid: 20374421
references.citation: Anesthesiology. 2007 Jul;107(1):91-8
references.pmid: 17585220
references.citation: Front Pharmacol. 2012 Mar 30;3:50
references.pmid: 22479247
references.citation: Biophys J. 2010 Jul 21;99(2):456-63
references.pmid: 20643063
references.citation: J Pharmacol Exp Ther. 2005 Feb;312(2):801-8
references.pmid: 15375177
references.citation: Br J Pharmacol. 2006 Jan;147 Suppl 1:S72-81
references.pmid: 16402123
references.citation: Biophys J. 2007 May 15;92(10):3513-23
references.pmid: 17325004
references.citation: Proc Natl Acad Sci U S A. 2012 Jan 10;109(2):E93-102
references.pmid: 22160714
references.citation: Proc Natl Acad Sci U S A. 2011 Nov 15;108(46):18825-30
references.pmid: 22042870
references.citation: J Pharmacol Exp Ther. 2005 Nov;315(2):517-25
references.pmid: 16033909
references.citation: Anesthesiology. 2009 Mar;110(3):582-90
references.pmid: 19225394
references.citation: Anesthesiology. 1996 May;84(5):1223-33; discussion 27A
references.pmid: 8624017
references.citation: Trends Pharmacol Sci. 2005 Oct;26(10):503-10
references.pmid: 16126282
references.citation: J Gen Physiol. 1981 Mar;77(3):233-53
references.pmid: 6265590
references.citation: Mol Pharmacol. 2003 Aug;64(2):373-81
references.pmid: 12869642
references.citation: Cell Mol Life Sci. 1999 Aug 15;55(10):1278-303
references.pmid: 10487207
references.citation: J Pharmacol Exp Ther. 2007 Sep;322(3):1076-83
references.pmid: 17569823
references.citation: Anesth Analg. 2007 Jan;104(1):84-91
references.pmid: 17179249
references.citation: Nat Rev Neurosci. 2008 May;9(5):370-86
references.pmid: 18425091
references.citation: Anesth Analg. 2008 Sep;107(3):875-8
references.pmid: 18713899
references.citation: Channels (Austin). 2010 Sep-Oct;4(5):422-8
references.pmid: 20935453
references.citation: Biophys J. 2011 Oct 5;101(7):1613-22
references.pmid: 21961587
references.citation: J Gen Physiol. 2004 Oct;124(4):349-56
references.pmid: 15365092
references.citation: Nat Commun. 2012;3:1102
references.pmid: 23033078
references.citation: Anesth Analg. 2008 Nov;107(5):1579-86
references.pmid: 18931215
references.citation: Structure. 2013 Aug 6;21(8):1307-16
references.pmid: 23891290
references.citation: Handb Exp Pharmacol. 2008;(182):3-29
references.pmid: 18175084
references.citation: J Anesth. 2011 Aug;25(4):609-13
references.pmid: 21656091
references.citation: Science. 1994 Sep 16;265(5179):1724-8
references.pmid: 8085162
references.citation: Br J Pharmacol. 2010 Feb;159(4):872-8
references.pmid: 20105175
references.citation: Biochim Biophys Acta. 2012 Sep;1818(9):2120-5
references.pmid: 22579978
references.citation: J Pharmacol Exp Ther. 2006 Jan;316(1):216-23
references.pmid: 16174800
references.citation: Neuropharmacology. 2011 Sep;61(4):699-706
references.pmid: 21651920
references.citation: Science. 2001 Dec 14;294(5550):2372-5
references.pmid: 11743207
references.citation: PLoS Comput Biol. 2013;9(6):e1003090
references.pmid: 23785267
references.citation: Anesthesiology. 2009 Sep;111(3):591-9
references.pmid: 19672182
references.citation: Biophys J. 2005 Jul;89(1):232-42
references.pmid: 15849254
references.citation: Biophys J. 2008 Jun;94(11):4260-9
references.pmid: 18310250
references.citation: Nature. 2012 May 20;486(7401):135-9
references.pmid: 22678296
references.citation: J Gen Physiol. 2012 Jun;139(6):507-16
references.pmid: 22641643