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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Jan 1;88(1):134–138. doi: 10.1073/pnas.88.1.134

Modulation of the general anesthetic sensitivity of a protein: a transition between two forms of firefly luciferase.

G W Moss 1, N P Franks 1, W R Lieb 1
PMCID: PMC50764  PMID: 1986359

Abstract

The activities of most proteins are relatively insensitive to general anesthetics. A notable exception is firefly luciferase, whose sensitivity to a wide range of anesthetic agents closely parallels that of whole animals. We have now found that this sensitivity can be controlled by ATP. The enzyme is insensitive at low (microM) concentrations of ATP and very sensitive at high (mM) concentrations. The differential sensitivity varies from anesthetic to anesthetic, being greatest (about a 100-fold difference) for molecules with large apolar segments. This suggests that anesthetic sensitivity is modulated by changes in the hydrophobicity of the anesthetic-binding pocket. Parallel changes in the binding of the substrate firefly luciferin, for which anesthetics compete, indicate that anesthetics bind at the same site as the luciferin substrate. These changes in the nature of the binding pocket modify not only the sensitivity to anesthetics but also the position of the "cutoff" in the homologous series of primary alcohol anesthetics; the cutoff position can vary from octanol to pentadecanol, depending upon the concentration of ATP. Our results suggest that particularly sensitive anesthetic target sites in the central nervous system may possess anesthetic-binding pockets whose polarities are regulated by neuromodulatory agents.

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Selected References

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

  1. Alifimoff J. K., Firestone L. L., Miller K. W. Anaesthetic potencies of primary alkanols: implications for the molecular dimensions of the anaesthetic site. Br J Pharmacol. 1989 Jan;96(1):9–16. doi: 10.1111/j.1476-5381.1989.tb11777.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Branchini B. R., Marschner T. M., Montemurro A. M. A convenient affinity chromatography-based purification of firefly luciferase. Anal Biochem. 1980 May 15;104(2):386–396. doi: 10.1016/0003-2697(80)90089-5. [DOI] [PubMed] [Google Scholar]
  3. Curry S., Lieb W. R., Franks N. P. Effects of general anesthetics on the bacterial luciferase enzyme from Vibrio harveyi: an anesthetic target site with differential sensitivity. Biochemistry. 1990 May 15;29(19):4641–4652. doi: 10.1021/bi00471a020. [DOI] [PubMed] [Google Scholar]
  4. DeLuca M., McElroy W. D. Kinetics of the firefly luciferase catalyzed reactions. Biochemistry. 1974 Feb 26;13(5):921–925. doi: 10.1021/bi00702a015. [DOI] [PubMed] [Google Scholar]
  5. Dorovska V. N., Varfolomeyev S. D., Kazanskaya N. F., Klyosov A. A., Martinek K. The influence of the geometric properties of the active centre on the specificity of -chymotrypsin catalysis. FEBS Lett. 1972 Jun 1;23(1):122–124. doi: 10.1016/0014-5793(72)80299-0. [DOI] [PubMed] [Google Scholar]
  6. Fastrez J., Fersht A. R. Mechanism of chymotrypsin. Structure, reactivity, and nonproductive binding relationships. Biochemistry. 1973 Mar 13;12(6):1067–1074. doi: 10.1021/bi00730a008. [DOI] [PubMed] [Google Scholar]
  7. Fersht A. R., Shindler J. S., Tsui W. C. Probing the limits of protein-amino acid side chain recognition with the aminoacyl-tRNA synthetases. Discrimination against phenylalanine by tyrosyl-tRNA synthetases. Biochemistry. 1980 Nov 25;19(24):5520–5524. doi: 10.1021/bi00565a009. [DOI] [PubMed] [Google Scholar]
  8. Franks N. P., Lieb W. R. Do general anaesthetics act by competitive binding to specific receptors? Nature. 1984 Aug 16;310(5978):599–601. doi: 10.1038/310599a0. [DOI] [PubMed] [Google Scholar]
  9. Franks N. P., Lieb W. R. Mapping of general anaesthetic target sites provides a molecular basis for cutoff effects. Nature. 1985 Jul 25;316(6026):349–351. doi: 10.1038/316349a0. [DOI] [PubMed] [Google Scholar]
  10. Franks N. P., Lieb W. R. Molecular mechanisms of general anaesthesia. Nature. 1982 Dec 9;300(5892):487–493. doi: 10.1038/300487a0. [DOI] [PubMed] [Google Scholar]
  11. Franks N. P., Lieb W. R. Partitioning of long-chain alcohols into lipid bilayers: implications for mechanisms of general anesthesia. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5116–5120. doi: 10.1073/pnas.83.14.5116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maze M. Transmembrane signalling and the holy grail of anesthesia. Anesthesiology. 1990 Jun;72(6):959–961. [PubMed] [Google Scholar]
  13. Miller K. W., Firestone L. L., Alifimoff J. K., Streicher P. Nonanesthetic alcohols dissolve in synaptic membranes without perturbing their lipids. Proc Natl Acad Sci U S A. 1989 Feb;86(3):1084–1087. doi: 10.1073/pnas.86.3.1084. [DOI] [PMC free article] [PubMed] [Google Scholar]

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