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Research ArticleNEUROPHARMACOLOGY

Multiple Specific Binding Targets for Inhaled Anesthetics in the Mammalian Brain

Maryellen Fazen Eckenhoff, Kin Chan and Roderic G. Eckenhoff
Journal of Pharmacology and Experimental Therapeutics January 2002, 300 (1) 172-179; DOI: https://doi.org/10.1124/jpet.300.1.172
Maryellen Fazen Eckenhoff
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Kin Chan
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Roderic G. Eckenhoff
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Abstract

Previous work showed widespread saturable binding of halothane in rat brain. To determine whether this represents selective binding to a few widespread proteins or less selective binding to many different proteins, we used [14C]halothane photolabeling and quantitative electrophoresis/autoradiography in rat cerebellar homogenates. Many proteins incorporate label. Stoichiometry values ranged from 0 to 4 at 0.2 mM [14C]halothane in a group of 24 randomly selected protein bands. Apparent IC50 values from unlabeled halothane competition experiments ranged from 0.2 to 2.0 mM, with soluble protein having significantly lower values (higher affinity) than membrane protein. Chloroform inhibited halothane labeling similar to unlabeled halothane but with higher apparent IC50 values, whereas isoflurane and an anesthetic, cyclobutane (1-chloro-1,2,2-trifluorocyclobutane), inhibited halothane labeling to a smaller degree. A nonanesthetic, cyclobutane (1,2-dichlorohexafluorocyclobutane), inhibited halothane labeling the least. We conclude that halothane binding motifs are sufficiently degenerate to be found in many proteins, both soluble and membrane-bound.

Footnotes

  • This work was supported by National Institute of General Medical Sciences Grants 51595 and 55876.

  • Abbreviations

    PBS
    phosphate-buffered saline
    PAGE
    polyacrylamide gel electrophoresis
    RD
    reflective densities
    OD
    optical densities
    PDB
    Protein Data Bank
    F3
    1-chloro-1,2,2-trifluorocyclobutane
    F6
    1,2-dichlorohexafluorocyclobutane
    • Received July 12, 2001.
    • Accepted October 9, 2001.
    • The American Society for Pharmacology and Experimental Therapeutics
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    Journal of Pharmacology and Experimental Therapeutics: 300 (1)
    Journal of Pharmacology and Experimental Therapeutics
    Vol. 300, Issue 1
    1 Jan 2002
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    Research ArticleNEUROPHARMACOLOGY

    Multiple Specific Binding Targets for Inhaled Anesthetics in the Mammalian Brain

    Maryellen Fazen Eckenhoff, Kin Chan and Roderic G. Eckenhoff
    Journal of Pharmacology and Experimental Therapeutics January 1, 2002, 300 (1) 172-179; DOI: https://doi.org/10.1124/jpet.300.1.172

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    Research ArticleNEUROPHARMACOLOGY

    Multiple Specific Binding Targets for Inhaled Anesthetics in the Mammalian Brain

    Maryellen Fazen Eckenhoff, Kin Chan and Roderic G. Eckenhoff
    Journal of Pharmacology and Experimental Therapeutics January 1, 2002, 300 (1) 172-179; DOI: https://doi.org/10.1124/jpet.300.1.172
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