Figure 5
From: A New Protein-Protein Interaction Sensor Based on Tripartite Split-GFP Association
Application of the tripartite split-GFP to study the ramapycin inducible FRB/FKBP interaction.
(a) GFP10 and GFP11 tags were fused to FRB and FKBP proteins. Rapamycin ligand binding brings both protein fusions into proximity, permitting GFP fluorescence reconstitution upon addition of GFP1–9. (b) Raw fluorescence progress curves for GFP1–9 complementation with soluble extracts of GFP10-FRB and FKBP-GFP11 fusions in presence (+RAP) or absence (−RAP) of rapamycin (starting time marked by addition of rapamycin to initiate complementation) (c) Fluorescence levels of GFP10-FRB and FKBP-GFP11 assayed at various concentrations (320, 160 and 40 nM in each) initiated by addition of rapamycin to 150 nM final concentration (black bar) and no rapamycin (gray bar). (d) Rapamycin dose curve (0.6 to 300 nM) for FRB/FKPB binding in vitro measured as final fluorescence after 1 h.
