Interaction in vitro; bait derived from adult head extract; prey produced and labeled by in vitro translation.

The inaD probe binds a ~130kDa band that is 1) recognized by anti-norpA antibody and 2) absent in norpA mutant background.

Radiolabelled inaD protein was overlayed onto blots containing adult head protein extracts from various genotypes separated by SDS-PAGE.

Source was membranes from adult heads of wild-type or mutant fly line; bait produced from endogenous gene; prey produced from endogenous gene.

trp, norpA and inaC, but not ninaE or Gαq, coimmunoprecipitate with inaD.

Interaction in vitro; bait derived from transfected COS cell lysate; prey produced as a recombinant fusion protein in bacterial system.

The consensus binding motif for inaD PDZ1 domain is X(F/Y/W)CF, as determined by peptide screening.

norpA was immunoprecipitated from transfected COS cells, separated by SDS-PAGE, transferred to nitrocellulose and incubated with GST-inaD.

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

GST-inaD was separated by SDS-PAGE, transferred to nitrocellulose and incubated with MBP-norpA.

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

Bipartite binding of inaD and norpA. The inaD PDZ1 domain binds the norpA C-terminus, and the inaD PDZ5 domain binds the norpA G box homology region.

Source was adult head membrane extract of wild-type fly line; bait produced from endogenous gene; prey produced from endogenous gene.

Using heat shock constructs in an inaD mutant background, the authors show that inaD interacts with norpA and inaC shortly after heat shock induced expression (3hr), long before these proteins reach the rhabdomeres and long before light

responses have recovered. This suggests that the complex is preassembled before transport.

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

The inaD PDZ1 domain forms a disulfide bridge with the norpA C-terminus, as evidenced by the fact that the complex persists during SDS-PAGE, but is disrupted by reducing agents.

Proteins were fractionated by size exclusion and complexes analyzed by SDS-PAGE in the presence or absence of reducing agents.

The strength of the norpA-inaD interaction decreases with mild acidifcation.

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

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

A mutant form of norpA in which 8 lysines are mutated to alanine (K880A,K884A,K887A,K888A,K891A,K898A,K899A,K902A) was used to determine the crystal structure.