Detection of protein-ligand interactions by NMR using reductive methylation of lysine residues

J Biomol NMR. 2008 Oct;42(2):143-8. doi: 10.1007/s10858-008-9274-y. Epub 2008 Sep 26.

Abstract

We show that reductive methylation of proteins can be used for highly sensitive NMR identification of conformational changes induced by metal- and small molecule binding, as well as protein-protein interactions. Reductive methylation of proteins introduces two (13)C-methyl groups on each lysine in the protein of interest. This method works well even when the lysines are not actively involved in the interaction, due to changes in the microenvironments of lysine residues. Most lysine residues are located on the protein exterior, and the exposed (13)C-methyl groups may exhibit rapid localized motions. These motions could be faster than the tumbling rate of the molecule as a whole. Thus, this technique has great potential in the study of large molecular weight systems which are currently beyond the scope of conventional NMR methods.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calmodulin / chemistry*
  • Humans
  • Ligands*
  • Lysine / chemistry*
  • Methylation
  • Nuclear Magnetic Resonance, Biomolecular / methods*

Substances

  • Calmodulin
  • Ligands
  • Lysine