Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey from embryo cDNA expression library).

Two-hybrid system: yeast LexA-BD/GAL4-AD. A constitutively active form of Rap1, Rap1[V12], was used as bait in Y2H screen.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast LexA-BD/GAL4-AD.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast LexA-BD/GAl4-AD

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

cno Ras association domains interact with constitutively active Rap1 stabilized in the active form with a non-hydrolyzable nucleotide GMPPNP. Complex formation determined by multi-angle light scattering with a molecular mass of 35.7 kDa for cno-Ras activation domain 1 and 30.4 kDa for cno-Ras activation domain 2.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in bacterial system.

cno Ras association domains interact with constitutively active Rap1 stabilized in the active form with a non-hydrolyzable nucleotide GMPPNP. Rap1-GMPPNP bound cno-Ras activation domain 1 with a Kd of 4.5 uM and bound cno-Ras activation domain 2 with a Kd of 8.7 uM.