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Mapping structural interactions using in-cell NMR spectroscopy (STINT-NMR).

David S Burz·Kaushik Dutta·David Cowburn·Alexander Shekhtman
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Abstract

We describe a high-throughput in-cell nuclear magnetic resonance (NMR)-based method for mapping the structural changes that accompany protein-protein interactions (STINT-NMR). The method entails sequentially expressing two (or more) proteins within a single bacterial cell in a time-controlled manner and monitoring the protein interactions using in-cell NMR spectroscopy. The resulting spectra provide a complete titration of the interaction and define structural details of the interacting surfaces at atomic resolution.

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Adaptor Proteins, Signal TransducingArabinoseAtaxin-3Binding SitesEndosomal Sorting Complexes Required for TransportEscherichia coliGene ExpressionIsopropyl ThiogalactosideModels, MolecularNerve Tissue ProteinsNuclear Magnetic Resonance, BiomolecularNuclear ProteinsPeptide FragmentsPhosphoproteinsPlasmidsProtein BindingProtein ConformationProtein Interaction MappingProtein Structure, QuaternaryProteinsRepressor ProteinsTransfectionUbiquitinJournal ArticleResearch Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov't

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David S Burz, Kaushik Dutta, David Cowburn, Alexander Shekhtman. 2006. Mapping structural interactions using in-cell NMR spectroscopy (STINT-NMR).. https://doi.org/10.1038/nmeth851

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