His3, His4, Caf1-55 complex reconstituted in vitro and subjected to analytical ultracentrifugation, interaction captured pairwise.

Interaction in vitro; peptides synthesized in vitro.

His3, His4, Caf1-55 complex reconstituted in vitro and subjected to gel filtration, interaction captured pairwise.

Interaction in vitro; peptides synthesized in vitro.

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

Proteins were separately purified, mixed and refolded. Complex formation was examined using gel filtration.

Positive control.

Recombinant histones were separately expressed and purified under denaturing conditions to directly solubilize inclusion bodies, then mixed (1mg each) and purified by size exclusion chromatography.

Recombinant His2A, His2B, His3 and His4 could be purified as a histone octamer complex.

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

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

Estimated mass of His3-His4 complex is consistent with a heterotetramer.