I-DIRT, a general method for distinguishing between specific and nonspecific protein interactions

J Proteome Res. Sep-Oct 2005;4(5):1752-6. doi: 10.1021/pr050225e.

Abstract

Isolation of protein complexes via affinity-tagged proteins provides a powerful tool for studying biological systems, but the technique is often compromised by co-enrichment of nonspecifically interacting proteins. We describe a new technique (I-DIRT) that distinguishes contaminants from bona fide interactors in immunopurifications, overcoming this most challenging problem in defining protein complexes. I-DIRT will be of broad value for studying protein complexes in biological systems that can be metabolically labeled.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Chromatography
  • DNA Polymerase II / chemistry
  • Fungal Proteins / chemistry
  • Lysine / chemistry
  • Macromolecular Substances / chemistry
  • Mass Spectrometry / methods*
  • Molecular Sequence Data
  • Open Reading Frames
  • Peptides / chemistry
  • Protein Binding
  • Protein Interaction Mapping*
  • Protein Structure, Tertiary
  • Proteins / chemistry*
  • Proteome*
  • Proteomics / methods*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Staphylococcal Protein A / chemistry
  • Trypsin / pharmacology

Substances

  • Fungal Proteins
  • Macromolecular Substances
  • Peptides
  • Proteins
  • Proteome
  • Saccharomyces cerevisiae Proteins
  • Staphylococcal Protein A
  • DNA Polymerase II
  • Trypsin
  • Lysine