Saturation transfer difference nuclear magnetic resonance spectroscopy as a method for screening proteins for anesthetic binding

John H. Streiff, Nenad O. Juranic, Slobodan I Macura, David Oman Warner, Keith A. Jones, William J. Perkins

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The effects of anesthetics on cellular function may result from direct interactions between anesthetic molecules and proteins. These interactions have a low affinity and are difficult to characterize. To identify proteins that bind anesthetics, we used nuclear magnetic resonance saturation transfer difference (STD) spectroscopy. The method is based on the nuclear Overhauser effect between bound anesthetic protons and all protein protons. To establish STD as a method for testing anesthetic binding to proteins, we conducted measurements on a series of protein/anesthetic solutions studied before by other methods. STD was able to identify that volatile anesthetics bind to bovine serum albumin, oleic acid reduces halothane binding to bovine serum albumin, and halothane binds to apomyoglobin but not lysozyme. Using STD, we found that halothane binding to calmodulin is Ca2+-dependent, which demonstrates anesthetic specificity for a protein conformation. Thus, STD is a powerful tool for investigating anesthetic-protein interactions because of its abilities to detect weak binding, to screen a single protein for binding of multiple anesthetics simultaneously, and to detect a change in anesthetic binding caused by conformational changes or competition with other ligands.

Original languageEnglish (US)
Pages (from-to)929-935
Number of pages7
JournalMolecular Pharmacology
Volume66
Issue number4
StatePublished - Oct 2004

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Protein Binding
Anesthetics
Magnetic Resonance Spectroscopy
Halothane
Bovine Serum Albumin
Proteins
Protons
Protein Conformation
Calmodulin
Oleic Acid
Muramidase
Spectrum Analysis
Carrier Proteins
Ligands

ASJC Scopus subject areas

  • Pharmacology

Cite this

Saturation transfer difference nuclear magnetic resonance spectroscopy as a method for screening proteins for anesthetic binding. / Streiff, John H.; Juranic, Nenad O.; Macura, Slobodan I; Warner, David Oman; Jones, Keith A.; Perkins, William J.

In: Molecular Pharmacology, Vol. 66, No. 4, 10.2004, p. 929-935.

Research output: Contribution to journalArticle

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