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.
Biochemistry
2014 Jan 14;531:135-42. doi: 10.1021/bi401492k.
Show Gene links
Show Anatomy links
Photoaffinity labeling the propofol binding site in GLIC.
Chiara DC
,
Gill JF
,
Chen Q
,
Tillman T
,
Dailey WP
,
Eckenhoff RG
,
Xu Y
,
Tang P
,
Cohen JB
.
???displayArticle.abstract???
Propofol, an intravenous general anesthetic, produces many of its anesthetic effects in vivo by potentiating the responses of GABA type A receptors (GABAAR), members of the superfamily of pentameric ligand-gated ion channels (pLGICs) that contain anion-selective channels. Propofol also inhibits pLGICs containing cation-selective channels, including nicotinic acetylcholine receptors and GLIC, a prokaryotic proton-gated homologue from Gloeobacter violaceus . In the structure of GLIC cocrystallized with propofol at pH 4 (presumed open/desensitized states), propofol was localized to an intrasubunit pocket at the extracellular end of the transmembrane domain within the bundle of transmembrane α-helices (Nury, H, et al. (2011) Nature 469, 428-431). To identify propofol binding sites in GLIC in solution, we used a recently developed photoreactive propofol analogue (2-isopropyl-5-[3-(trifluoromethyl)-3H-diazirin-3-yl]phenol or AziPm) that acts as an anesthetic in vivo and potentiates GABAAR in vitro. For GLIC expressed in Xenopus oocytes, propofol and AziPm inhibited current responses at pH 5.5 (EC20) with IC50 values of 20 and 50 μM, respectively. When [(3)H]AziPm (7 μM) was used to photolabel detergent-solubilized, affinity-purified GLIC at pH 4.4, protein microsequencing identified propofol-inhibitable photolabeling of three residues in the GLIC transmembrane domain: Met-205, Tyr-254, and Asn-307 in the M1, M3, and M4 transmembrane helices, respectively. Thus, for GLIC in solution, propofol and AziPm bind competitively to a site in proximity to these residues, which, in the GLIC crystal structure, are in contact with the propofol bound in the intrasubunit pocket.
Arevalo,
Gating-enhanced accessibility of hydrophobic sites within the transmembrane region of the nicotinic acetylcholine receptor's {delta}-subunit. A time-resolved photolabeling study.
2005, Pubmed
Arevalo,
Gating-enhanced accessibility of hydrophobic sites within the transmembrane region of the nicotinic acetylcholine receptor's {delta}-subunit. A time-resolved photolabeling study.
2005,
Pubmed
Baenziger,
3D structure and allosteric modulation of the transmembrane domain of pentameric ligand-gated ion channels.
2011,
Pubmed
Bertaccini,
Assessment of homology templates and an anesthetic binding site within the γ-aminobutyric acid receptor.
2013,
Pubmed
Bocquet,
A prokaryotic proton-gated ion channel from the nicotinic acetylcholine receptor family.
2007,
Pubmed
,
Xenbase
Bondarenko,
NMR structures of the transmembrane domains of the α4β2 nAChR.
2012,
Pubmed
Brauer,
Use of o-phthalaldehyde to reduce background during automated Edman degradation.
1984,
Pubmed
Chen,
Anesthetic binding in a pentameric ligand-gated ion channel: GLIC.
2010,
Pubmed
Chiara,
Specificity of intersubunit general anesthetic-binding sites in the transmembrane domain of the human α1β3γ2 γ-aminobutyric acid type A (GABAA) receptor.
2013,
Pubmed
Chiara,
Identification of nicotinic acetylcholine receptor amino acids photolabeled by the volatile anesthetic halothane.
2003,
Pubmed
Chiara,
Mapping general anesthetic binding site(s) in human α1β3 γ-aminobutyric acid type A receptors with [³H]TDBzl-etomidate, a photoreactive etomidate analogue.
2012,
Pubmed
Chiara,
Time-resolved photolabeling of the nicotinic acetylcholine receptor by [3H]azietomidate, an open-state inhibitor.
2009,
Pubmed
,
Xenbase
Crimmins,
In situ chemical cleavage of proteins immobilized to glass-fiber and polyvinylidenedifluoride membranes: cleavage at tryptophan residues with 2-(2'-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine to obtain internal amino acid sequence.
1990,
Pubmed
Dellisanti,
Site-directed spin labeling reveals pentameric ligand-gated ion channel gating motions.
2013,
Pubmed
,
Xenbase
Dilger,
Evidence for direct actions of general anesthetics on an ion channel protein. A new look at a unified mechanism of action.
1994,
Pubmed
Flood,
Alpha 4 beta 2 neuronal nicotinic acetylcholine receptors in the central nervous system are inhibited by isoflurane and propofol, but alpha 7-type nicotinic acetylcholine receptors are unaffected.
1997,
Pubmed
,
Xenbase
Franks,
General anaesthesia: from molecular targets to neuronal pathways of sleep and arousal.
2008,
Pubmed
Garcia,
[3H]Benzophenone photolabeling identifies state-dependent changes in nicotinic acetylcholine receptor structure.
2007,
Pubmed
,
Xenbase
Ghosh,
Propofol binding to the resting state of the gloeobacter violaceus ligand-gated ion channel (GLIC) induces structural changes in the inter- and intrasubunit transmembrane domain (TMD) cavities.
2013,
Pubmed
,
Xenbase
Hall,
m-Azipropofol (AziPm) a photoactive analogue of the intravenous general anesthetic propofol.
2010,
Pubmed
,
Xenbase
Hamouda,
Multiple transmembrane binding sites for p-trifluoromethyldiazirinyl-etomidate, a photoreactive Torpedo nicotinic acetylcholine receptor allosteric inhibitor.
2011,
Pubmed
Hamouda,
Probing the structure of the affinity-purified and lipid-reconstituted torpedo nicotinic acetylcholine receptor.
2008,
Pubmed
Hemmings,
Emerging molecular mechanisms of general anesthetic action.
2005,
Pubmed
Jayakar,
Identification of propofol binding sites in a nicotinic acetylcholine receptor with a photoreactive propofol analog.
2013,
Pubmed
Krasowski,
General anesthetic potencies of a series of propofol analogs correlate with potency for potentiation of gamma-aminobutyric acid (GABA) current at the GABA(A) receptor but not with lipid solubility.
2001,
Pubmed
,
Xenbase
Li,
Identification of a GABAA receptor anesthetic binding site at subunit interfaces by photolabeling with an etomidate analog.
2006,
Pubmed
Middleton,
Mapping of the acetylcholine binding site of the nicotinic acetylcholine receptor: [3H]nicotine as an agonist photoaffinity label.
1991,
Pubmed
Mowrey,
Asymmetric ligand binding facilitates conformational transitions in pentameric ligand-gated ion channels.
2013,
Pubmed
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
Rudolph,
Molecular and neuronal substrates for general anaesthetics.
2004,
Pubmed
Sauguet,
Structural basis for potentiation by alcohols and anaesthetics in a ligand-gated ion channel.
2013,
Pubmed
Spurny,
Multisite binding of a general anesthetic to the prokaryotic pentameric Erwinia chrysanthemi ligand-gated ion channel (ELIC).
2013,
Pubmed
,
Xenbase
Stewart,
p-(4-Azipentyl)propofol: a potent photoreactive general anesthetic derivative of propofol.
2011,
Pubmed
,
Xenbase
Unwin,
Refined structure of the nicotinic acetylcholine receptor at 4A resolution.
2005,
Pubmed
Velisetty,
Desensitization mechanism in prokaryotic ligand-gated ion channel.
2012,
Pubmed
Velisetty,
Conformational transitions underlying pore opening and desensitization in membrane-embedded Gloeobacter violaceus ligand-gated ion channel (GLIC).
2012,
Pubmed
Violet,
Differential sensitivities of mammalian neuronal and muscle nicotinic acetylcholine receptors to general anesthetics.
1997,
Pubmed
,
Xenbase
Vodovozova,
Photoaffinity labeling and its application in structural biology.
2007,
Pubmed
Weng,
Anesthetic sensitivity of the Gloeobacter violaceus proton-gated ion channel.
2010,
Pubmed
,
Xenbase
Yamodo,
Conformational changes in the nicotinic acetylcholine receptor during gating and desensitization.
2010,
Pubmed
Yip,
A propofol binding site on mammalian GABAA receptors identified by photolabeling.
2013,
Pubmed
