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

Syx1A, Snap24, nSyb form trimeric complex. Although all 3 proteins are required for complex formation, interactions are captured pairwise.

nSyb bearing v-SNARE vesicles contain a lipid-bound fluorescence resonance energy transfer donor-acceptor pair. When v-SNARE vesicles fuse to t-SNARE vesicles (bearing Syx1A and Snap25), the donor-acceptor pair is diluted, resulting in dequenching of the fluorescence signal.

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

Source was adult heads of wild-type or comt⁴ temperature sensitive fly line; bait produced from endogenous gene; prey produced from endogenous gene.

Immunoprecipitation of nSyb was more efficient in comt⁴ temperature sensitive mutant background than in wild-type flies.

Flies were exposed to 38oC for 10min for tissue collection and preparation of head homogenates.