A novel NMR method for determining the interfaces of large protein-protein complexes

Nat Struct Biol. 2000 Mar;7(3):220-3. doi: 10.1038/73331.

Abstract

Identification of the interfaces of large (Mr > 50,000) protein-protein complexes in solution by high resolution NMR has typically been achieved using experiments involving chemical shift perturbation and/or hydrogen-deuterium exchange of the main chain amide groups of the proteins. Interfaces identified using these techniques, however, are not always identical to those revealed using X-ray crystallography. In order to identify the contact residues in a large protein-protein complex more accurately, we developed a novel NMR method that uses cross-saturation phenomena in combination with TROSY detection in an optimally deuterium labeled system.

Publication types

  • Comparative Study

MeSH terms

  • Binding Sites
  • Crystallography, X-Ray
  • Deuterium / metabolism
  • Diffusion
  • Humans
  • Immunoglobulin Fc Fragments / chemistry*
  • Immunoglobulin Fc Fragments / metabolism*
  • Isotope Labeling
  • Macromolecular Substances
  • Models, Molecular
  • Molecular Weight
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Protein Binding
  • Protein Structure, Secondary
  • Protons
  • Reproducibility of Results
  • Solutions
  • Staphylococcal Protein A / chemistry*
  • Staphylococcal Protein A / metabolism*
  • Staphylococcus aureus

Substances

  • Immunoglobulin Fc Fragments
  • Macromolecular Substances
  • Protons
  • Solutions
  • Staphylococcal Protein A
  • Deuterium