An NMR Method to Pinpoint Supramolecular Ligand Binding to Basic Residues on Proteins.
Related Articles An NMR Method to Pinpoint Supramolecular Ligand Binding to Basic Residues on Proteins.
Angew Chem Int Ed Engl. 2017 Sep 06;:
Authors: Hogeweg A, Sowislok A, Schrader T, Beuck C
Abstract
Targeting protein surfaces involved in protein-protein interactions using supramolecular chemistry is a rapidly growing field. NMR spectroscopy is the method of choice to map ligand binding sites with single residue resolution by amide chemical shift perturbation and line broadening. However, large aromatic ligands affect NMR signals over a greater distance, and the binding site cannot be determined unambiguously by relying on backbone signals only. We employed Lys- and Arg-specific H2(C)N NMR experiments to directly observe the side chain atoms in close contact with the ligand, for which the most drastic changes in the NMR signals are expected. Binding of lysine- and arginine-specific supramolecular tweezers to the small hPin1 WW domain was studied as a model system. The H2(C)N spectra track the terminal CH2 groups of all two Lys and four Arg residues, revealing significant differences in their binding kinetics and allow to clearly pinpoint the order of tweezers binding.
PMID: 28877391 [PubMed - as supplied by publisher]
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