A highly sensitive protein-protein interaction assay based on Gaussia luciferase.
Protein-fragment complementation assays (PCAs) provide a general strategy to study the dynamics of protein-protein interactions in vivo and in vitro. The full potential of PCA requires assays that are fully reversible and sensitive at subendogenous protein expression levels. We describe a new assay that meets these criteria, based on the Gaussia princeps luciferase enzyme, demonstrating chemical reversal, and induction and inhibition of a key interaction linking insulin and TGFbeta signaling.
Remy
Ingrid
I
Département de Biochimie, Université de Montréal, C.P. 6128, Succursale centre-ville, Montréal, Québec, H3C 3J7, Canada.
Michnick
Stephen W
SW
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D033342:Copepoda / Q000201:enzymology*; D008156:Luciferases / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D008164:Luminescent Proteins / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D008164:Luminescent Proteins; D010446:Peptide Fragments; D008156:Luciferases