Protein-protein interaction site mapping using NMR-detected mutational scanning

J Biomol NMR. 2007 Jun;38(2):133-7. doi: 10.1007/s10858-007-9154-x. Epub 2007 Apr 20.

Abstract

We demonstrate a novel NMR method for the mapping of protein-protein interaction sites. In our approach protein-protein binding sites are mapped by competition binding experiments using indirect NMR reporter technology and Ala positional scanning. The methodology provides high sensitivity, ease of implementation and high-throughput capabilities. The feasibility of the technique is demonstrated with an application to the beta-Catenin/Tcf4 complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding, Competitive
  • Humans
  • Ligands
  • Models, Molecular
  • Mutation / genetics*
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Protein Interaction Mapping / methods*
  • Reproducibility of Results
  • TCF Transcription Factors / metabolism
  • Transcription Factor 7-Like 2 Protein
  • beta Catenin / metabolism

Substances

  • Ligands
  • TCF Transcription Factors
  • TCF7L2 protein, human
  • Transcription Factor 7-Like 2 Protein
  • beta Catenin