(15)N and (13)C group-selective techniques extend the scope of STD NMR detection of weak host-guest interactions and ligand screening

Chembiochem. 2010 Oct 18;11(15):2182-7. doi: 10.1002/cbic.201000317.

Abstract

Saturation transfer difference (STD) is a valuable tool for studying the binding of small molecules to large biomolecules and for obtaining detailed information on the binding epitopes. Here, we demonstrate that the proposed (15)N/(13)C variants of group-selective, "GS-STD" experiments provide a powerful approach to mapping the binding epitope of a ligand even in the absence of efficient spin diffusion within the target protein. Therefore, these experimental variants broaden the scope of STD studies to smaller and/or more-dynamic targets. The STD spectra obtained in four different experimental setups (selective (1)H STD, (15)N GS-STD, (13)C(Ar) and (13)C(aliphatic) GS-STD approaches) revealed that the signal-intensity pattern of the difference spectra is affected by both the type and the spatial distribution of the excited "transmitter" atoms, as well as by the efficiency of the spin-diffusion-mediated magnetization transfer. The performance of the experiments is demonstrated on a system by using the lectin, galectin-1 and its carbohydrate ligand, lactose.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Isotopes / chemistry
  • Dimerization
  • Galectin 1 / chemistry
  • Lactose / chemistry
  • Ligands*
  • Nitrogen Isotopes / chemistry
  • Nuclear Magnetic Resonance, Biomolecular*
  • Protein Binding
  • Protein Structure, Tertiary

Substances

  • Carbon Isotopes
  • Galectin 1
  • Ligands
  • Nitrogen Isotopes
  • Lactose