I-DIRT, a general method for distinguishing between specific and nonspecific protein interactions.
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.
Tackett
Alan J
AJ
The Rockefeller University, New York, NY 10021, USA.
DeGrasse
Jeffrey A
JA
Sekedat
Matthew D
MD
Oeffinger
Marlene
M
Rout
Michael P
MP
Chait
Brian T
BT
D000595:Amino Acid Sequence; D002845:Chromatography; D004257:DNA Polymerase II / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D046911:Macromolecular Substances / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry; D014357:Trypsin / Q000494:pharmacology
D005656:Fungal Proteins; D046911:Macromolecular Substances; D010455:Peptides; D011506:Proteins; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins; D013205:Staphylococcal Protein A; D004257:DNA Polymerase II; D014357:Trypsin; D008239:Lysine