A modular cross-linking approach for exploring protein interactions.
A method is described for the elucidation of protein-protein interactions using novel cross-linking reagents and mass spectrometry. The method incorporates (1) a modular solid-phase synthetic strategy for generating the cross-linking reagents, (2) enrichment and digestion of cross-linked proteins using microconcentrators, (3) mass spectrometric analysis of cross-linked peptides, and (4) comprehensive computational analysis of the cross-linking data. This integrated approach has been applied to the study of cross-linking between the components of the heterodimeric protein complex negative cofactor 2.
Trester-Zedlitz
Michelle
M
Laboratories of Synthetic Protein Chemistry, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Kamada
Katsuhiko
K
Burley
Stephen K
SK
Fenyö
David
D
Chait
Brian T
BT
Muir
Tom W
TW
D000180:Acrylic Resins / Q000737:chemistry; D000595:Amino Acid Sequence; D001710:Biotin / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D005998:Glycine / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014157:Transcription Factors / Q000737:chemistry*; D000099166:Polyacrylamides
D000180:Acrylic Resins; D001710:Biotin; D003432:Cross-Linking Reagents; D005998:Glycine; D010446:Peptide Fragments; D010750:Phosphoproteins; D011092:Polyethylene Glycols; D013438:Sulfhydryl Compounds; D014157:Transcription Factors; D000099166:Polyacrylamides; C075995:down-regulator of transcription 1