Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011 Jun;174(3):434-42.
doi: 10.1016/j.jsb.2011.04.001. Epub 2011 Apr 9.

Rapid Identification of Protein-Protein Interfaces for the Construction of a Complex Model Based on Multiple Unassigned Signals by Using Time-Sharing NMR Measurements

Affiliations

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 et al. J Struct Biol. .

Abstract

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 (15)N-labeled amino acid and several amino acids (13)C-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 ω(1)-TROSY-type backbone (1)H-(15)N and aromatic (1)H-(13)C shift correlations together with single quantum methyl (1)H-(13)C 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.

Similar articles

See all similar articles

Cited by 1 article

  • The quiet renaissance of protein nuclear magnetic resonance.
    Barrett PJ, Chen J, Cho MK, Kim JH, Lu Z, Mathew S, Peng D, Song Y, Van Horn WD, Zhuang T, Sönnichsen FD, Sanders CR. Barrett PJ, et al. Biochemistry. 2013 Feb 26;52(8):1303-20. doi: 10.1021/bi4000436. Epub 2013 Feb 12. Biochemistry. 2013. PMID: 23368985 Free PMC article. Review.

Publication types

MeSH terms

LinkOut - more resources

Feedback