The strength and specificity of protein complex formation is crucial for most life processes and is determined by interactions between residues in the binding partners. Double-mutant cycle analysis provides a strategy for studying the energetic coupling between amino acids at the interfaces of such complexes. Here we show that these pairwise interaction energies can be determined from a single high-resolution native mass spectrum by measuring the intensities of the complexes formed by the two wild-type proteins, the complex of each wild-type protein with a mutant protein, and the complex of the two mutant proteins. This native mass spectrometry approach, which obviates the need for error-prone measurements of binding constants, can provide information regarding multiple interactions in a single spectrum much like nuclear Overhauser effects (NOEs) in nuclear magnetic resonance. Importantly, our results show that specific inter-protein contacts in solution are maintained in the gas phase.
CITATION STYLE
Sokolovski, M., Cveticanin, J., Hayoun, D., Korobko, I., Sharon, M., & Horovitz, A. (2017). Measuring inter-protein pairwise interaction energies from a single native mass spectrum by double-mutant cycle analysis. Nature Communications, 8(1). https://doi.org/10.1038/s41467-017-00285-1
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