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 GAL4-BD/GAL4-AD
Q70L mutation locks protein in GTP-bound activated state
Bait expressed in stably transfected cells.
Prey expressed in transfected cells.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey derived from wild-type embryonic cytosolic extract.
Interaction in vitro; bait produced as a recombinant fusion protein in S2 cells; prey derived from recombinant fusion protein in S2 cells.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey derived from S2 cell extract (endogenous gene).
Specificity S-score=13.08.
Constitutively active (GTP-locked) form of Rab was used for pull down. Parallel pull downs performed for several Rabs proteins in this study. Yield was measured using spectral counts. A specificity score (S score) was calculated, where greater weight was given to interactions seen with fewer baits.
Y380S, coordinates relative to Rip11-PB
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced from tagged transfected construct.
