Figure 2.

Determination of the Orc1 domain responsible for binding AlF-C2. (A) The wild-type (lane 1) and various mutants of Orc1 (lanes 2–6), represented in (B), were translated in vitro as T7-tagged proteins and incubated with glutathione beads coupled with GST–AlF-C2. Proteins bound to the beads were separated by SDS–PAGE and blotted with anti-T7 antibody (upper panel). The 1/10 volume of unbound fractions was also analyzed (lower panel). (B) Schematic representation of Orc1 and its truncation mutants. The BAH domain, ATPase (AAA+ family of ATPase) homology domain and AlF-C2 binding domain are shown with amino acid residue number.