Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Methods Enzymol
2018 Jan 01;602:369-389. doi: 10.1016/bs.mie.2018.01.014.
Show Gene links
Show Anatomy links
Combining Mutations and Electrophysiology to Map Anesthetic Sites on Ligand-Gated Ion Channels.
Forman SA
.
???displayArticle.abstract???
General anesthetics are known to act in part by binding to and altering the function of pentameric ligand-gated ion channels such as nicotinic acetylcholine and γ-aminobutyric acid type A receptors. Combining heterologous expression of the subunits that assemble to form these ion channels, mutagenesis techniques and voltage-clamp electrophysiology have enabled a variety of "structure-function" approaches to questions of where anesthetic binds to these ion channels and how they enhance or inhibit channel function. Here, we review the evolution of concepts and experimental strategies during the last three decades, since molecular biological and electrophysiological tools became widely used. Topics covered include: (1) structural models as interpretive frameworks, (2) various electrophysiological approaches and their limitations, (3) Monod-Wyman-Changeux allosteric models as functional frameworks, (4) structural strategies including chimeras and point mutations, and (5) methods based on cysteine substitution and covalent modification. We discuss in particular depth the experimental design considerations for substituted cysteine modification-protection studies.
Akabas,
Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the alpha subunit.
1994, Pubmed,
Xenbase
Akabas,
Identification of acetylcholine receptor channel-lining residues in the entire M2 segment of the alpha subunit.
1994,
Pubmed
,
Xenbase
Akk,
Neurosteroid access to the GABAA receptor.
2005,
Pubmed
Bali,
GABA-induced intersubunit conformational movement in the GABAA receptor alpha 1M1-beta 2M3 transmembrane subunit interface: experimental basis for homology modeling of an intravenous anesthetic binding site.
2009,
Pubmed
,
Xenbase
Bali,
Defining the propofol binding site location on the GABAA receptor.
2004,
Pubmed
,
Xenbase
Belelli,
The interaction of the general anesthetic etomidate with the gamma-aminobutyric acid type A receptor is influenced by a single amino acid.
1997,
Pubmed
,
Xenbase
Borghese,
GABA(A) receptor transmembrane amino acids are critical for alcohol action: disulfide cross-linking and alkyl methanethiosulfonate labeling reveal relative location of binding sites.
2014,
Pubmed
,
Xenbase
Brett,
A method for the rapid exchange of solutions bathing excised membrane patches.
1986,
Pubmed
Colquhoun,
Binding, gating, affinity and efficacy: the interpretation of structure-activity relationships for agonists and of the effects of mutating receptors.
1998,
Pubmed
Desai,
Gamma-amino butyric acid type A receptor mutations at beta2N265 alter etomidate efficacy while preserving basal and agonist-dependent activity.
2009,
Pubmed
,
Xenbase
Dilger,
The effects of isoflurane on acetylcholine receptor channels.: 2. Currents elicited by rapid perfusion of acetylcholine.
1993,
Pubmed
Dilger,
Evidence for direct actions of general anesthetics on an ion channel protein. A new look at a unified mechanism of action.
1994,
Pubmed
Dilger,
Effects of isoflurane on acetylcholine receptor channels. 1. Single-channel currents.
1992,
Pubmed
Feng,
Etomidate produces similar allosteric modulation in α1β3δ and α1β3γ2L GABA(A) receptors.
2014,
Pubmed
,
Xenbase
Feng,
Barbiturates require the N terminus and first transmembrane domain of the delta subunit for enhancement of alpha1beta3delta GABAA receptor currents.
2010,
Pubmed
Forman,
A discrete site for general anesthetics on a postsynaptic receptor.
1995,
Pubmed
,
Xenbase
Forman,
Novel modulation of a nicotinic receptor channel mutant reveals that the open state is stabilized by ethanol.
1999,
Pubmed
,
Xenbase
Forman,
Homologous mutations on different subunits cause unequal but additive effects on n-alcohol block in the nicotinic receptor pore.
1997,
Pubmed
,
Xenbase
Forman,
Mapping General Anesthetic Sites in Heteromeric γ-Aminobutyric Acid Type A Receptors Reveals a Potential For Targeting Receptor Subtypes.
2016,
Pubmed
Forman,
A hydrophobic photolabel inhibits nicotinic acetylcholine receptors via open-channel block following a slow step.
1999,
Pubmed
Fourati,
Barbiturates Bind in the GLIC Ion Channel Pore and Cause Inhibition by Stabilizing a Closed State.
2017,
Pubmed
,
Xenbase
Goren,
Loose protein packing around the extracellular half of the GABA(A) receptor beta1 subunit M2 channel-lining segment.
2004,
Pubmed
,
Xenbase
Harrison,
Positive modulation of human gamma-aminobutyric acid type A and glycine receptors by the inhalation anesthetic isoflurane.
1993,
Pubmed
Hill-Venning,
Subunit-dependent interaction of the general anaesthetic etomidate with the gamma-aminobutyric acid type A receptor.
1997,
Pubmed
,
Xenbase
Jansen,
State-dependent cross-linking of the M2 and M3 segments: functional basis for the alignment of GABAA and acetylcholine receptor M3 segments.
2006,
Pubmed
,
Xenbase
Jenkins,
Tryptophan scanning mutagenesis in TM4 of the GABA(A) receptor alpha1 subunit: implications for modulation by inhaled anesthetics and ion channel structure.
2002,
Pubmed
Jurd,
General anesthetic actions in vivo strongly attenuated by a point mutation in the GABA(A) receptor beta3 subunit.
2003,
Pubmed
Koltchine,
Agonist gating and isoflurane potentiation in the human gamma-aminobutyric acid type A receptor determined by the volume of a second transmembrane domain residue.
1999,
Pubmed
Krasowski,
Methionine 286 in transmembrane domain 3 of the GABAA receptor beta subunit controls a binding cavity for propofol and other alkylphenol general anesthetics.
2001,
Pubmed
Krasowski,
Propofol and other intravenous anesthetics have sites of action on the gamma-aminobutyric acid type A receptor distinct from that for isoflurane.
1998,
Pubmed
Laurent,
Sites of Anesthetic Inhibitory Action on a Cationic Ligand-Gated Ion Channel.
2016,
Pubmed
Leonard,
Evidence that the M2 membrane-spanning region lines the ion channel pore of the nicotinic receptor.
1988,
Pubmed
,
Xenbase
Lewis,
The ion channel properties of a rat recombinant neuronal nicotinic receptor are dependent on the host cell type.
1997,
Pubmed
,
Xenbase
Li,
Identification of a GABAA receptor anesthetic binding site at subunit interfaces by photolabeling with an etomidate analog.
2006,
Pubmed
Maldifassi,
Functional sites involved in modulation of the GABAA receptor channel by the intravenous anesthetics propofol, etomidate and pentobarbital.
2016,
Pubmed
,
Xenbase
Mihic,
Sites of alcohol and volatile anaesthetic action on GABA(A) and glycine receptors.
1997,
Pubmed
,
Xenbase
Miller,
General anesthetic and specific effects of ethanol on acetylcholine receptors.
1987,
Pubmed
Nourmahnad,
Tryptophan and Cysteine Mutations in M1 Helices of α1β3γ2L γ-Aminobutyric Acid Type A Receptors Indicate Distinct Intersubunit Sites for Four Intravenous Anesthetics and One Orphan Site.
2016,
Pubmed
,
Xenbase
Nury,
X-ray structures of general anaesthetics bound to a pentameric ligand-gated ion channel.
2011,
Pubmed
Pan,
Structure of the pentameric ligand-gated ion channel GLIC bound with anesthetic ketamine.
2012,
Pubmed
,
Xenbase
Ruesch,
An allosteric coagonist model for propofol effects on α1β2γ2L γ-aminobutyric acid type A receptors.
2012,
Pubmed
,
Xenbase
Rüsch,
Gating allosterism at a single class of etomidate sites on alpha1beta2gamma2L GABA A receptors accounts for both direct activation and agonist modulation.
2004,
Pubmed
,
Xenbase
Serafini,
Structural domains of the human GABAA receptor 3 subunit involved in the actions of pentobarbital.
2000,
Pubmed
Shu,
Characteristics of concatemeric GABA(A) receptors containing α4/δ subunits expressed in Xenopus oocytes.
2012,
Pubmed
,
Xenbase
Spurny,
Multisite binding of a general anesthetic to the prokaryotic pentameric Erwinia chrysanthemi ligand-gated ion channel (ELIC).
2013,
Pubmed
,
Xenbase
Stern,
A Cysteine Substitution Probes β3H267 Interactions with Propofol and Other Potent Anesthetics in α1β3γ2L γ-Aminobutyric Acid Type A Receptors.
2016,
Pubmed
,
Xenbase
Stewart,
Tryptophan mutations at azi-etomidate photo-incorporation sites on alpha1 or beta2 subunits enhance GABAA receptor gating and reduce etomidate modulation.
2008,
Pubmed
,
Xenbase
Stewart,
Cysteine substitutions define etomidate binding and gating linkages in the α-M1 domain of γ-aminobutyric acid type A (GABAA) receptors.
2013,
Pubmed
,
Xenbase
Stewart,
Mutations at beta N265 in γ-aminobutyric acid type A receptors alter both binding affinity and efficacy of potent anesthetics.
2014,
Pubmed
,
Xenbase
Stewart,
State-dependent etomidate occupancy of its allosteric agonist sites measured in a cysteine-substituted GABAA receptor.
2013,
Pubmed
,
Xenbase
Stühmer,
Electrophysiological recording from Xenopus oocytes.
1992,
Pubmed
,
Xenbase
Ueno,
Tryptophan scanning mutagenesis in TM2 of the GABA(A) receptor alpha subunit: effects on channel gating and regulation by ethanol.
2000,
Pubmed
,
Xenbase
Williams,
Gamma-aminobutyric acid increases the water accessibility of M3 membrane-spanning segment residues in gamma-aminobutyric acid type A receptors.
1999,
Pubmed
Xu,
Identification of channel-lining residues in the M2 membrane-spanning segment of the GABA(A) receptor alpha1 subunit.
1996,
Pubmed
,
Xenbase
Zhou,
The n-alcohol site in the nicotinic receptor pore is a hydrophobic patch.
2000,
Pubmed
Ziemba,
Correction for Inhibition Leads to an Allosteric Co-Agonist Model for Pentobarbital Modulation and Activation of α1β3γ2L GABAA Receptors.
2016,
Pubmed
,
Xenbase
