Using oxidative crosslinking and proximity labeling to quantitatively characterize protein-protein and protein-Peptide complexes.
The quantitative analysis of protein-protein and protein-peptide complexes is of fundamental importance in biochemistry. We report here that nickel-catalyzed proximity biotinylation and Ru(II)(bpy)(3)(2+)-mediated oxidative crosslinking can be used to measure the equilibrium dissociation constant and stoichiometry of protein complexes. Only small amounts of protein are required, neither of the binding partners must be immobilized on a surface, and no special instrumentation is necessary. This chemistry should provide a useful complement to existing methods for the analysis of protein-protein and protein-peptide interactions.
Amini
Frank
F
Center for Biomedical Inventions, Departments of Internal Medicine and Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Denison
Carilee
C
Lin
Hai Jui
HJ
Kuo
Lillian
L
Kodadek
Thomas
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D001710:Biotin / Q000737:chemistry; D002384:Catalysis; D017931:DNA Primers; D008969:Molecular Sequence Data; D009532:Nickel / Q000737:chemistry; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*
D017931:DNA Primers; D010455:Peptides; D011506:Proteins; D001710:Biotin; D009532:Nickel