Figure 2. Development of AGS8 peptide.

(A) Schematic diagram of AGS8-C and the AGS8-peptides (CP1 and CP9) which inhibited AGS8-Gβγ interaction. The AGS8-peptides were developed based on amino acid sequences of the Gβγ interaction domain of AGS8. CP1 and CP2 represented A¹⁴⁹⁴– V¹⁵²² and S¹⁵⁰⁸– I¹⁵³⁷ of rat AGS8 (ABB82299) respectively. (B) The example of GST-pulldown assay of GST-AGS8-C with Gβ₁γ₂. GST-fusion protein (100 nM) was incubated with recombinant human Gβ₁γ₂ (10 nM) in the presence of AGS8-peptides (CP1 or CP9). Proteins were then adsorbed to a glutathione matrix and retained G-protein subunits identified by immunoblotting following gel electrophoresis. The representative of 6 independent experiments with similar results. (C) The densitometric analysis of GST-pulldown assay of GST-AGS8-C with Gβ₁γ₂ in the presence of AGS8-peptides. n = 6 with peptides of ∼97% HPLC purity.