Source was adult heads of wild-type fly line; bait produced from endogenous gene; prey produced from endogenous gene.
"F3" construct
K1259A, "PDZ*" construct, coordinates corresponding to Spn-PG
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; Spn produced as a recombinant fusion protein in bacterial system; Nrx-1 peptide chemically synthesized.
Interaction in vitro; bait chemically synthesized; prey produced as a recombinant fusion protein in bacterial system.
The Spn PDZ domain binds the Nrx-1 PDZ binding motif peptide with Kd = 50uM (ten-fold lower affinity than interaction of the RhoGAP100F PDZ domain with the Nrx-1 peptide).
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in bacterial system.