Periodate-triggered cross-linking of DOPA-containing peptide-protein complexes.
Chemical cross-linking is a powerful methodology for analyzing proteins-small molecule and protein-protein interactions. We describe the development of a new chemical cross-linking reaction for the study of protein complexes. Specifically, we show that molecules containing an ortho dihydroxyarene unit can be oxidized selectively with sodium periodate in the presence of native proteins, producing an ortho quinone intermediate that can cross-link with suitable nearby protein residues. We demonstrate the efficacy and specificity of this chemistry for a peptide-protein complex and also deduce the binding site of an artificial activation domain on a proteasome subcomplex.
Burdine
Lyle
L
Center for Biomedical Inventions University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-8573, USA.
Gillette
Thomas G
TG
Lin
Hai-Jun
HJ
Kodadek
Thomas
T
D000595:Amino Acid Sequence; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004295:Dihydroxyphenylalanine / Q000737:chemistry*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry*; D010504:Periodic Acid / Q000737:chemistry*; D011809:Quinones / Q000737:chemistry; D012097:Repressor Proteins / Q000032:analysis / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000737:chemistry*
D003432:Cross-Linking Reagents; C055002:GAL80 protein, S cerevisiae; D010455:Peptides; D011809:Quinones; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D010504:Periodic Acid; D004295:Dihydroxyphenylalanine; C009288:metaperiodate