Rapid identification of protein-protein interfaces for the construction of a complex model based on multiple unassigned signals by using time-sharing NMR measurements

Yuya Kodama, Michael L. Reese, Nobuhisa Shimba, Katsuki Ono, Eiji Kanamori, Volker Dötsch, Shuji Noguchi, Yoshifumi Fukunishi, Ei ichiro Suzuki, Ichio Shimada, Hideo Takahashi

Research output: Contribution to journalArticle

3 Scopus citations


Protein-protein interactions are necessary for various cellular processes, and therefore, information related to protein-protein interactions and structural information of complexes is invaluable. To identify protein-protein interfaces using NMR, resonance assignments are generally necessary to analyze the data; however, they are time consuming to collect, especially for large proteins. In this paper, we present a rapid, effective, and unbiased approach for the identification of a protein-protein interface without resonance assignments. This approach requires only a single set of 2D titration experiments of a single protein sample, labeled with a unique combination of an 15N-labeled amino acid and several amino acids 13C-labeled on specific atoms. To rapidly obtain high resolution data, we applied a new pulse sequence for time-shared NMR measurements that allowed simultaneous detection of a ω1-TROSY-type backbone 1H-15N and aromatic 1H-13C shift correlations together with single quantum methyl 1H-13C shift correlations. We developed a structure-based computational approach, that uses our experimental data to search the protein surfaces in an unbiased manner to identify the residues involved in the protein-protein interface. Finally, we demonstrated that the obtained information of the molecular interface could be directly leveraged to support protein-protein docking studies. Such rapid construction of a complex model provides valuable information and enables more efficient biochemical characterization of a protein-protein complex, for instance, as the first step in structure-guided drug development.

Original languageEnglish (US)
Pages (from-to)434-442
Number of pages9
JournalJournal of Structural Biology
Issue number3
Publication statusPublished - Jun 2011



  • Assignment free
  • NMR
  • Protein-protein docking
  • Protein-protein interaction
  • Time-sharing measurements

ASJC Scopus subject areas

  • Structural Biology

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