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Organic solvents identify specific ligand binding sites on protein surfaces

Nature Biotechnology volume 15, pages 264–268 (1997)Cite this article

Abstract

Enzymes frequently recognize substrates and pharmaceutical drugs through specific binding interactions in deep pockets on the protein surface. We show how the specificity-determining substrate binding site of hen egg-white lysozyme (HEWL) can be readily identified in aqueous solution by nuclear magnetic resonance spectroscopy using small organic solvent molecules as detection probes. Exchange of magnetization between the 1H nuclei of the protein and the ligands through dipole-dipole interactions is observed which allows the modeling of their position and orientation at the binding site. Combined with site-specific binding constants measured by titration experiments with different organic solvents, the method can provide important information for rational drug design. In addition, the lifetime of nonspecific interactions of HEWL with organic solvents is shown to be in the sub-nanosecond time range.

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Author notes

  1. Gottfried Otting: Corresponding author (e-mail: go@mfn.ki.se).

Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Karolinska Institute, 171 77, Stockholm, Sweden

    Edvards Liepinsh

Authors
  1. Edvards Liepinsh
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  2. Gottfried Otting
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Liepinsh, E., Otting, G. Organic solvents identify specific ligand binding sites on protein surfaces. Nat Biotechnol 15, 264–268 (1997). https://doi.org/10.1038/nbt0397-264

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