Speeding-up the Determination of Protein-Ligand Affinities by STD NMR: The Reduced Data Set STD NMR Approach (rd-STD NMR)

Anal Chem. 2024 Jan 16;96(2):615-619. doi: 10.1021/acs.analchem.3c03980. Epub 2024 Jan 2.

Abstract

STD NMR spectroscopy is a powerful ligand-observed NMR tool for screening and characterizing the interactions of small molecules and low molecular weight fragments with a given macromolecule, identifying the main intermolecular contacts in the bound state. It is also a powerful analytical technique for the accurate determination of protein-ligand dissociation constants (KD) of medium-to-weak affinity, of interest in the pharmaceutical industry. However, accurate KD determination and epitope mapping requires a long series of experiments at increasing saturation times to carry out a full analysis using the so-called STD NMR build-up curve approach and apply the "initial slopes approximation". Here, we have developed a new protocol to bypass this important limitation, which allows us to obtain initial slopes by using just two saturation times and, hence, to very quickly determine precise protein-ligand dissociation constants by STD NMR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Epitope Mapping
  • Ligands
  • Magnetic Resonance Imaging*
  • Magnetic Resonance Spectroscopy / methods
  • Protein Binding
  • Proteins* / chemistry

Substances

  • Ligands
  • Proteins