Figure 5
From: Unraveling the molecular architecture of a G protein-coupled receptor/β-arrestin/Erk module complex
Experimental validation of the predicted β-arrestin-Raf1 and β-arrestin-ERK2 interaction sites.
A) Equal amounts of GST-Raf1-RBD fusion protein were incubated with 25 ng of β-arrestin 1 and with or without 0.5mM of the indicated peptide. B) Representative blot for β-arrestin-Raf1 interaction site sequentially probed with anti-β-arrestin and anti-GST antibodies. B) Equal amounts of GST-ERK2 fusion protein were incubated with 25 ng of β-arrestin 1 and with or without 0.5mM of the indicated peptide. Representative blot for β-arrestin-ERK2 interaction site sequentially probed with anti-β-arrestin and anti-ERK antibodies. Immunoblots were quantified using Odyssey software and data were expressed as the ratio of β-arrestin 1 over GST-Raf1-RBD or GST-ERK respectively. The maximum response measured with the Control peptide was chosen as 100%. Data represent means ± SEM from three independent experiments. *,**: p < 0.05 specific peptides versus control peptides.
