Organic solvents identify specific ligand binding sites on protein surfaces
- 1 March 1997
- journal article
- Published by Springer Nature in Nature Biotechnology
- Vol. 15 (3), 264-268
- https://doi.org/10.1038/nbt0397-264
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.Keywords
This publication has 24 references indexed in Scilit:
- Locating and characterizing binding sites on proteinsNature Biotechnology, 1996
- An Experimental Approach to Mapping the Binding Surfaces of Crystalline ProteinsThe Journal of Physical Chemistry, 1996
- Dissection of Protein–Carbohydrate Interactions in Mutant Hen Egg-white Lysozyme Complexes and their Hydrolytic ActivityJournal of Molecular Biology, 1995
- Crystal Structure of the Mutant D52S Hen Egg White Lysozyme with an Oligosaccharide ProductJournal of Molecular Biology, 1994
- The solvent dependence of enzyme specificityBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1994
- 1H Nuclear Magnetic Resonance Studies of Hen Lysozyme-N-Acetylglucosamine Oligosaccharide Complexes in Solution: Application of Chemical Shifts for the Comparison of Conformational Changes in Solution and in the CrystalJournal of Molecular Biology, 1994
- Refinement of an enzyme complex with inhibitor bound at partial occupancyJournal of Molecular Biology, 1992
- Study of ethanol-lysozyme interactions using neutron diffractionBiochemistry, 1985
- X-ray crystallography of the binding of the bacterial cell wall trisaccharide NAM-NAG-NAM to lysozymeNature, 1979
- Structural analysis of denaturant-protein interactions: Comparison between the effects of bromoethanol and SDS on denaturation and renaturation of triclinic lysozymeEuropean Biophysics Journal, 1978