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

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...

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Veröffentlicht in:	Nature methods 2006-02, Vol.3 (2), p.91-93
Hauptverfasser:	Shekhtman, Alexander, Burz, David S, Dutta, Kaushik, Cowburn, David
Format:	Artikel
Sprache:	eng
Schlagworte:	Adaptor Proteins, Signal Transducing - chemistry Adaptor Proteins, Signal Transducing - genetics Adaptor Proteins, Signal Transducing - metabolism Arabinose - pharmacology Ataxin-3 Binding Sites Bioinformatics Biological Microscopy Biological Techniques Biomedical and Life Sciences Biomedical Engineering/Biotechnology brief-communication Endosomal Sorting Complexes Required for Transport Escherichia coli - genetics Escherichia coli - metabolism Gene Expression - drug effects Isopropyl Thiogalactoside - pharmacology Life Sciences Measurement Methods Models, Molecular Nerve Tissue Proteins - chemistry Nerve Tissue Proteins - genetics Nerve Tissue Proteins - metabolism NMR Nuclear magnetic resonance Nuclear magnetic resonance spectroscopy Nuclear Magnetic Resonance, Biomolecular - methods Nuclear Proteins Peptide Fragments - chemistry Peptide Fragments - genetics Peptide Fragments - metabolism Phosphoproteins - chemistry Phosphoproteins - genetics Phosphoproteins - metabolism Physiological aspects Plasmids - genetics Protein Binding Protein Conformation Protein Interaction Mapping - methods Protein Structure, Quaternary Protein-protein interactions Proteins - chemistry Proteins - genetics Proteins - metabolism Proteomics Repressor Proteins Spectroscopy Transfection Ubiquitin - chemistry Ubiquitin - genetics Ubiquitin - metabolism
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Beschreibung
Zusammenfassung:	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.
ISSN:	1548-7091 1548-7105
DOI:	10.1038/nmeth851