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

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Complex modes of BicD homodimerization were characterized. The C-terminal domain (CTD) and the central region, each capable of homodimerizing in this assay, are also able to interact with each other.

Intramolecular interactions within BicD are thought hold the protein in an inactive conformation in the absence of cargo. Association of a cargo-binding protein with the BicD CTD presumably competes out the self-interaction, freeing the N-terminus of BicD to stimulate cargo transport.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

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

Recombinant BicD forms a homodimer.

Two-hybrid system: yeast GAL4-BD/GAL4-AD

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

The N-terminus of BicD (fused to GAL4 activation domain) interacts with the C-terminal region of BicD (fused to GAL4 DNA-binding domain).

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

BicD forms a highly asymmetric parallel coiled coil dimer; one chain is very straight, with the second chain wrapped around it in a highly supercoiled state.

EM consistent with a BicD dimer.

Interaction in vitro; protein produced as a recombinant fusion protein in baculovirus and Sf9 cell system.

BicD forms a homodimer in crystallo. Crystal structure of a F684I mutant form of BicD was determined. While this mutant still formed a homodimer, it was found to be in a different coiled-coil register than wild-type BicD.

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

Two-hybrid system: yeast GAL4-BD/GAL4-AD; positive control.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).