Fluorescence polarization assay to quantify protein-protein interactions.
A fluorescence polarization assay can be used to evaluate the strength of a protein-protein interaction. A green fluorescent protein variant is fused to one of the protein partners. The formation of a complex is then deduced from an increase in fluorescence polarization, and the equilibrium dissociation constant of the complex is determined in a homogeneous aqueous environment. The assay is demonstrated by using the interaction of the S-protein and S-peptide fragments of ribonuclease A as a case study.
Park
Sang-Hyun
SH
Department of Cellular and Molecular Pharmacology, University of California-San Francisco, San Francisco, CA, USA.
Raines
Ronald T
RT
D000818:Animals; D005454:Fluorescence Polarization / Q000379:methods*; D049452:Green Fluorescent Proteins; D007700:Kinetics; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000378:metabolism
D008164:Luminescent Proteins; D010446:Peptide Fragments; D011506:Proteins; D049452:Green Fluorescent Proteins; C074799:ribonuclease A (21-124); D012259:Ribonuclease, Pancreatic