Source was embryos of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

A two step coIP of Sumo was utilized (His followed by Flag).

Drosophila Sumo enzyme cycle components expressed in a bacterial system with bait and prey.

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

Source was cell extract of S2R+ cell line; bait produced from transfected construct; prey produced from transfected construct.

smt3 biotinylated in vivo by coexpression of bacterial BirA enzyme. Proteins purified under denaturing conditions to inactivate enzymes and dissociate smt3-binding proteins, leaving only covalently bound proteins.