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

Nat Methods. 2006 Feb;3(2):91-3. doi: 10.1038/nmeth851.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / chemistry
  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism
  • Arabinose / pharmacology
  • Ataxin-3
  • Binding Sites
  • Endosomal Sorting Complexes Required for Transport
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression / drug effects
  • Isopropyl Thiogalactoside / pharmacology
  • Models, Molecular
  • Nerve Tissue Proteins / chemistry
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Nuclear Proteins
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Phosphoproteins / chemistry
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Plasmids / genetics
  • Protein Binding
  • Protein Conformation
  • Protein Interaction Mapping / methods*
  • Protein Structure, Quaternary*
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / metabolism
  • Repressor Proteins
  • Transfection
  • Ubiquitin / chemistry
  • Ubiquitin / genetics
  • Ubiquitin / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • Endosomal Sorting Complexes Required for Transport
  • Nerve Tissue Proteins
  • Nuclear Proteins
  • Peptide Fragments
  • Phosphoproteins
  • Proteins
  • Repressor Proteins
  • STAM2 protein, human
  • Ubiquitin
  • Isopropyl Thiogalactoside
  • Arabinose
  • ATXN3 protein, human
  • Ataxin-3