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 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.
Burz
David S
DS
State University of New York at Albany, Department of Chemistry, 1400 Washington Ave., Albany, New York 12222, USA.
Dutta
Kaushik
K
Cowburn
David
D
Shekhtman
Alexander
A
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001089:Arabinose / Q000494:pharmacology; D000067699:Ataxin-3; D001665:Binding Sites; D056827:Endosomal Sorting Complexes Required for Transport; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008958:Models, Molecular; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009687:Nuclear Proteins; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping / Q000379:methods*; D020836:Protein Structure, Quaternary; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins; D014162:Transfection; D025801:Ubiquitin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D056827:Endosomal Sorting Complexes Required for Transport; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D011506:Proteins; D012097:Repressor Proteins; C491647:STAM2 protein, human; D025801:Ubiquitin; D007544:Isopropyl Thiogalactoside; D001089:Arabinose; C092341:ATXN3 protein, human; D000067699:Ataxin-3