Related ArticlesNMR study of volatile anesthetic binding to nicotinic acetylcholine receptors.
Biophys J. 2000 Feb;78(2):746-51
Authors: Xu Y, Seto T, Tang P, Firestone L
New lines of evidence suggest that volatile anesthetics interact specifically with proteins. Direct binding analysis, however, has been largely limited to soluble proteins. In this study, specific interaction was investigated between isoflurane, a clinically important volatile anesthetic, and membrane-bound nicotinic acetylcholine receptors (nAChRs) from Torpedo electroplax, using (19)F nuclear magnetic resonance spectroscopy and gas chromatography. The receptors were reconstituted into 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) lipid vesicles. After correcting for nonspecific partitioning into the lipid, the equilibrium dissociation constant, K(d), of isoflurane binding to nAChR at 15 degrees C was found to be 0.36 +/- 0.03 mM. This value is within the clinically relevant concentration range of the agent. Based on the receptor concentrations in the vesicle suspension assayed by the bicinchoninic acid method and the fraction of bound isoflurane, X(b), determined by gas chromatography, an estimate of an average of 9-10 specifically bound isoflurane molecules can be made for each receptor, or two for each subunit. Upon binding, the transverse relaxation time constant (T(2)) of (19)F resonance of isoflurane is decreased by nearly three orders of magnitude, indicating a dramatic reduction in the mobility of specifically bound isoflurane. Kinetic analysis reveals that the off rate of binding, k(-1), is 1.7 x 10(4) s(-1). The on rate, k(+1), can thus be calculated to be approximately 4.8 x 10(7) M(-1) s(-1), suggesting a nearly diffusion-limited association. This is in contrast to anesthetic binding to a soluble protein, bovine serum albumin (BSA), where k(+1) and k(-1) are at least an order of magnitude slower. It is concluded that the presence of lipids may be critical for the correct evaluation of binding kinetics between volatile anesthetics and neuronal receptors.
Neurotoxin II Bound to Acetylcholine Receptors in Native Membranes Studied by Dynamic Nuclear Polarization NMR
Neurotoxin II Bound to Acetylcholine Receptors in Native Membranes Studied by Dynamic Nuclear Polarization NMR
Arne H. Linden, Sascha Lange, W. Trent Franks, U?mit Akbey, Edgar Specker, Barth-Jan van Rossum and Hartmut Oschkinat
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja206999c/aop/images/medium/ja-2011-06999c_0003.gif
Journal of the American Chemical Society
DOI: 10.1021/ja206999c
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/4d1LYPHtCFw
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Nmr structure and action on nicotinic acetylcholine receptors of water-soluble domain of human lynx1.
NMR STRUCTURE AND ACTION ON NICOTINIC ACETYLCHOLINE RECEPTORS OF WATER-SOLUBLE DOMAIN OF HUMAN LYNX1.
NMR STRUCTURE AND ACTION ON NICOTINIC ACETYLCHOLINE RECEPTORS OF WATER-SOLUBLE DOMAIN OF HUMAN LYNX1.
J Biol Chem. 2011 Jan 20;
Authors: Lyukmanova EN, Shenkarev ZO, Shulepko MA, Mineev KS, D'Hoedt D, Kasheverov IE, Filkin SY, Krivolapova AP, Janickova H, Dolezal V, Dolgikh DA, Arseniev AS, Bertrand D, Tsetlin VI, Kirpichnikov MP
Discovery of proteins expressed in the central nervous system sharing the three-finger structure with snake...
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[NMR paper] Application of the 19F NMR technique to observe binding of the general anesthetic hal
Application of the 19F NMR technique to observe binding of the general anesthetic halothane to human serum albumin.
Related Articles Application of the 19F NMR technique to observe binding of the general anesthetic halothane to human serum albumin.
Anal Sci. 2004 Oct;20(10):1475-7
Authors: Shikii K, Sakurai S, Utsumi H, Seki H, Tashiro M
19F NMR techniques were employed to characterize the binding property of the widely used general anesthetic halothane with human serum albumin (HSA). It was found that 19F(1H) NOE and 2D 1H-19F HOESY...
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11-24-2010 10:01 PM
[NMR paper] Towards structure determination of neurotoxin II bound to nicotinic acetylcholine rec
Towards structure determination of neurotoxin II bound to nicotinic acetylcholine receptor: a solid-state NMR approach.
Related Articles Towards structure determination of neurotoxin II bound to nicotinic acetylcholine receptor: a solid-state NMR approach.
FEBS Lett. 2004 Apr 30;564(3):319-24
Authors: Krabben L, van Rossum BJ, Castellani F, Bocharov E, Schulga AA, Arseniev AS, Weise C, Hucho F, Oschkinat H
Solid-state magic-angle spinning nuclear magnetic resonance (NMR) has sufficient resolving power for full assignment of resonances and...
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[NMR paper] Peptide hormone binding to G-protein-coupled receptors: structural characterization v
Peptide hormone binding to G-protein-coupled receptors: structural characterization via NMR techniques.
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Med Res Rev. 2001 Sep;21(5):450-71
Authors: Mierke DF, Giragossian C
G-protein-coupled receptors (GPCRs) allow cells to respond to calcium, hormones, and neurotransmitters. Not surprisingly, they currently make up the largest family of validated drug targets. Rational drug design for molecular regulators targeting GPCRs...
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11-19-2010 08:44 PM
[NMR paper] Probing the agonist binding pocket in the nicotinic acetylcholine receptor: a high-re
Probing the agonist binding pocket in the nicotinic acetylcholine receptor: a high-resolution solid-state NMR approach.
Related Articles Probing the agonist binding pocket in the nicotinic acetylcholine receptor: a high-resolution solid-state NMR approach.
Biochemistry. 1998 Jul 28;37(30):10854-9
Authors: Williamson PT, Gröbner G, Spooner PJ, Miller KW, Watts A
Acetylcholine, the agonist for the nicotinic acetylcholine receptor, has been observed directly when bound specifically to its binding site in the fully functional receptor-enriched...
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11-17-2010 11:15 PM
[NMR paper] Volatile anesthetics compete for common binding sites on bovine serum albumin: a 19F-
Volatile anesthetics compete for common binding sites on bovine serum albumin: a 19F-NMR study.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Volatile anesthetics compete for common binding sites on bovine serum albumin: a 19F-NMR study.
Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6478-82
Authors: Dubois BW, Cherian SF, Evers AS
There is controversy as to the molecular nature of volatile anesthetic target sites. One proposal is that volatile anesthetics...
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08-22-2010 03:01 AM
[NMR paper] 19F-NMR spin-spin relaxation (T2) method for characterizing volatile anesthetic bindi
19F-NMR spin-spin relaxation (T2) method for characterizing volatile anesthetic binding to proteins. Analysis of isoflurane binding to serum albumin.
Related Articles 19F-NMR spin-spin relaxation (T2) method for characterizing volatile anesthetic binding to proteins. Analysis of isoflurane binding to serum albumin.
Biochemistry. 1992 Aug 11;31(31):7069-76
Authors: Dubois BW, Evers AS
This paper characterizes the low-affinity ligand binding interactions of a fluorinated volatile anesthetic, isoflurane (CHF2OCHClCF3), with bovine serum albumin...