FIGURE 4.

Role of Tyr⁵¹⁰ in association of Dbl with substrate GTPases. GST fusion proteins of the indicated RhoA (A) and Cdc42 (B) variants were overexpressed in E. coli, purified on glutathione affinity chromatography, and their nucleotide content manipulated as described under “Materials and Methods.” Top panels, GEF presence in affinity precipitates. Middle panels, Dbl expression in respective cell lysates. Lower panels, GST-GTPases “baits” were visualized by Ponceau Red staining of the blotted membranes. Data are representative of three independent experiments. IB, immunoblot.