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. 2002 Sep;83(3):1413–1420. doi: 10.1016/S0006-3495(02)73912-X

Effects of volatile anesthetic on channel structure of gramicidin A.

Pei Tang 1, Pravat K Mandal 1, Martha Zegarra 1
PMCID: PMC1302240  PMID: 12202367

Abstract

Volatile anesthetic agent, 1-chloro-1,2,2-trifluorocyclobutane (F3), was found to alter gramicidin A channel function by enhancing Na(+) transport (. Biophys. J. 77:739-746). Whether this functional change is associated with structural alternation is evaluated by circular dichroism and nuclear magnetic resonance spectroscopy. The circular dichroism and nuclear magnetic resonance results indicate that at low millimolar concentrations, 1-chloro-1,2,2-trifluorocyclobutane causes minimal changes in gramicidin A channel structure in sodium dodecyl sulfate micelles. All hydrogen bonds between channel backbones are well maintained in the presence of 1-chloro-1,2,2-trifluorocyclobutane, and the channel structure is stable. The finding supports the notion that low affinity drugs such as volatile anesthetics and alcohols can cause significant changes in protein function without necessarily producing associated changes in protein structure. To understand the molecular mechanism of general anesthesia, it is important to recognize that in addition to structural changes, other protein properties, including dynamic characteristics of channel motions, may also be of functional significance.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abdul-Manan N., Hinton J. F. Conformation states of gramicidin A along the pathway to the formation of channels in model membranes determined by 2D NMR and circular dichroism spectroscopy. Biochemistry. 1994 Jun 7;33(22):6773–6783. doi: 10.1021/bi00188a005. [DOI] [PubMed] [Google Scholar]
  2. Arseniev A. S., Barsukov I. L., Bystrov V. F., Lomize A. L., Ovchinnikov YuA 1H-NMR study of gramicidin A transmembrane ion channel. Head-to-head right-handed, single-stranded helices. FEBS Lett. 1985 Jul 8;186(2):168–174. doi: 10.1016/0014-5793(85)80702-x. [DOI] [PubMed] [Google Scholar]
  3. Bax A., Vuister G. W., Grzesiek S., Delaglio F., Wang A. C., Tschudin R., Zhu G. Measurement of homo- and heteronuclear J couplings from quantitative J correlation. Methods Enzymol. 1994;239:79–105. doi: 10.1016/s0076-6879(94)39004-5. [DOI] [PubMed] [Google Scholar]
  4. Becker M. D., Greathouse D. V., Koeppe R. E., 2nd, Andersen O. S. Amino acid sequence modulation of gramicidin channel function: effects of tryptophan-to-phenylalanine substitutions on the single-channel conductance and duration. Biochemistry. 1991 Sep 10;30(36):8830–8839. doi: 10.1021/bi00100a015. [DOI] [PubMed] [Google Scholar]
  5. Bhattacharya A. A., Curry S., Franks N. P. Binding of the general anesthetics propofol and halothane to human serum albumin. High resolution crystal structures. J Biol Chem. 2000 Dec 8;275(49):38731–38738. doi: 10.1074/jbc.M005460200. [DOI] [PubMed] [Google Scholar]
  6. Carter D. C., He X. M. Structure of human serum albumin. Science. 1990 Jul 20;249(4966):302–303. doi: 10.1126/science.2374930. [DOI] [PubMed] [Google Scholar]
  7. Clore G. M., Gronenborn A. M. New methods of structure refinement for macromolecular structure determination by NMR. Proc Natl Acad Sci U S A. 1998 May 26;95(11):5891–5898. doi: 10.1073/pnas.95.11.5891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Conti E., Franks N. P., Brick P. Crystal structure of firefly luciferase throws light on a superfamily of adenylate-forming enzymes. Structure. 1996 Mar 15;4(3):287–298. doi: 10.1016/s0969-2126(96)00033-0. [DOI] [PubMed] [Google Scholar]
  9. Cross T. A. Solid-state nuclear magnetic resonance characterization of gramicidin channel structure. Methods Enzymol. 1997;289:672–696. doi: 10.1016/s0076-6879(97)89070-2. [DOI] [PubMed] [Google Scholar]
  10. Delaglio F., Grzesiek S., Vuister G. W., Zhu G., Pfeifer J., Bax A. NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR. 1995 Nov;6(3):277–293. doi: 10.1007/BF00197809. [DOI] [PubMed] [Google Scholar]
  11. Eckenhoff R. G., Johansson J. S. Molecular interactions between inhaled anesthetics and proteins. Pharmacol Rev. 1997 Dec;49(4):343–367. [PubMed] [Google Scholar]
  12. Eckenhoff R. G., Petersen C. E., Ha C. E., Bhagavan N. V. Inhaled anesthetic binding sites in human serum albumin. J Biol Chem. 2000 Sep 29;275(39):30439–30444. doi: 10.1074/jbc.M005052200. [DOI] [PubMed] [Google Scholar]
  13. Eckenhoff R. G., Tanner J. W. Differential halothane binding and effects on serum albumin and myoglobin. Biophys J. 1998 Jul;75(1):477–483. doi: 10.1016/S0006-3495(98)77536-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Forman S. A., Miller K. W., Yellen G. A discrete site for general anesthetics on a postsynaptic receptor. Mol Pharmacol. 1995 Oct;48(4):574–581. [PubMed] [Google Scholar]
  15. Franks N. P., Jenkins A., Conti E., Lieb W. R., Brick P. Structural basis for the inhibition of firefly luciferase by a general anesthetic. Biophys J. 1998 Nov;75(5):2205–2211. doi: 10.1016/S0006-3495(98)77664-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Franks N. P., Lieb W. R. Molecular and cellular mechanisms of general anaesthesia. Nature. 1994 Feb 17;367(6464):607–614. doi: 10.1038/367607a0. [DOI] [PubMed] [Google Scholar]
  17. Greathouse D. V., Hinton J. F., Kim K. S., Koeppe R. E., 2nd Gramicidin A/short-chain phospholipid dispersions: chain length dependence of gramicidin conformation and lipid organization. Biochemistry. 1994 Apr 12;33(14):4291–4299. doi: 10.1021/bi00180a025. [DOI] [PubMed] [Google Scholar]
  18. Johansson J. S., Zou H., Tanner J. W. Bound volatile general anesthetics alter both local protein dynamics and global protein stability. Anesthesiology. 1999 Jan;90(1):235–245. doi: 10.1097/00000542-199901000-00030. [DOI] [PubMed] [Google Scholar]
  19. Ketchem R. R., Hu W., Cross T. A. High-resolution conformation of gramicidin A in a lipid bilayer by solid-state NMR. Science. 1993 Sep 10;261(5127):1457–1460. doi: 10.1126/science.7690158. [DOI] [PubMed] [Google Scholar]
  20. Ketchem R., Roux B., Cross T. High-resolution polypeptide structure in a lamellar phase lipid environment from solid state NMR derived orientational constraints. Structure. 1997 Dec 15;5(12):1655–1669. doi: 10.1016/s0969-2126(97)00312-2. [DOI] [PubMed] [Google Scholar]
  21. Marion D., Wüthrich K. Application of phase sensitive two-dimensional correlated spectroscopy (COSY) for measurements of 1H-1H spin-spin coupling constants in proteins. Biochem Biophys Res Commun. 1983 Jun 29;113(3):967–974. doi: 10.1016/0006-291x(83)91093-8. [DOI] [PubMed] [Google Scholar]
  22. Masotti L., Spisni A., Urry D. W. Conformational studies on the gramicidin A transmembrane channel in lipid micelles and liposomes. Cell Biophys. 1980 Sep;2(3):241–251. doi: 10.1007/BF02790452. [DOI] [PubMed] [Google Scholar]
  23. Mihic S. J., Ye Q., Wick M. J., Koltchine V. V., Krasowski M. D., Finn S. E., Mascia M. P., Valenzuela C. F., Hanson K. K., Greenblatt E. P. Sites of alcohol and volatile anaesthetic action on GABA(A) and glycine receptors. Nature. 1997 Sep 25;389(6649):385–389. doi: 10.1038/38738. [DOI] [PubMed] [Google Scholar]
  24. Nilges M., Clore G. M., Gronenborn A. M. Determination of three-dimensional structures of proteins from interproton distance data by dynamical simulated annealing from a random array of atoms. Circumventing problems associated with folding. FEBS Lett. 1988 Oct 24;239(1):129–136. doi: 10.1016/0014-5793(88)80559-3. [DOI] [PubMed] [Google Scholar]
  25. North C., Cafiso D. S. Contrasting membrane localization and behavior of halogenated cyclobutanes that follow or violate the Meyer-Overton hypothesis of general anesthetic potency. Biophys J. 1997 Apr;72(4):1754–1761. doi: 10.1016/S0006-3495(97)78821-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Piotto M., Saudek V., Sklenár V. Gradient-tailored excitation for single-quantum NMR spectroscopy of aqueous solutions. J Biomol NMR. 1992 Nov;2(6):661–665. doi: 10.1007/BF02192855. [DOI] [PubMed] [Google Scholar]
  27. Tang P., Eckenhoff R. G., Xu Y. General anesthetic binding to gramicidin A: the structural requirements. Biophys J. 2000 Apr;78(4):1804–1809. doi: 10.1016/S0006-3495(00)76730-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tang P., Hu J., Liachenko S., Xu Y. Distinctly different interactions of anesthetic and nonimmobilizer with transmembrane channel peptides. Biophys J. 1999 Aug;77(2):739–746. doi: 10.1016/S0006-3495(99)76928-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tang P., Simplaceanu V., Xu Y. Structural consequences of anesthetic and nonimmobilizer interaction with gramicidin A channels. Biophys J. 1999 May;76(5):2346–2350. doi: 10.1016/S0006-3495(99)77391-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tang P., Yan B., Xu Y. Different distribution of fluorinated anesthetics and nonanesthetics in model membrane: a 19F NMR study. Biophys J. 1997 Apr;72(4):1676–1682. doi: 10.1016/S0006-3495(97)78813-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Townsley L. E., Tucker W. A., Sham S., Hinton J. F. Structures of gramicidins A, B, and C incorporated into sodium dodecyl sulfate micelles. Biochemistry. 2001 Oct 2;40(39):11676–11686. doi: 10.1021/bi010942w. [DOI] [PubMed] [Google Scholar]
  32. Veatch W. R., Fossel E. T., Blout E. R. The conformation of gramicidin A. Biochemistry. 1974 Dec 17;13(26):5249–5256. doi: 10.1021/bi00723a001. [DOI] [PubMed] [Google Scholar]
  33. Wallace B. A. Structure of gramicidin A. Biophys J. 1986 Jan;49(1):295–306. doi: 10.1016/S0006-3495(86)83642-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wallace B. A., Veatch W. R., Blout E. R. Conformation of gramicidin A in phospholipid vesicles: circular dichroism studies of effects of ion binding, chemical modification, and lipid structure. Biochemistry. 1981 Sep 29;20(20):5754–5760. doi: 10.1021/bi00523a018. [DOI] [PubMed] [Google Scholar]
  35. Xu Y., Seto T., Tang P., Firestone L. NMR study of volatile anesthetic binding to nicotinic acetylcholine receptors. Biophys J. 2000 Feb;78(2):746–751. doi: 10.1016/S0006-3495(00)76632-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Xu Y., Tang P. Amphiphilic sites for general anesthetic action? Evidence from 129Xe-[1H] intermolecular nuclear Overhauser effects. Biochim Biophys Acta. 1997 Jan 14;1323(1):154–162. doi: 10.1016/s0005-2736(96)00184-8. [DOI] [PubMed] [Google Scholar]

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