Two-hybrid system: yeast GAL4-BD/GAL4-AD
Bait Rala protein carried the constitutively activating mutation G20V, as well as a C198S mutation to prevent membrane attachment via geranylgeranylation.
Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey from embryo cDNA expression library).
coordinates relative to 139kDa isoform (GEFmeso-PG)
Interaction was tested with constitutively active (G20V) and dominant-negative (S25N) versions of Rala.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.
