Fig. 2.

Reconstitution of recombinant AMC and identification of the conserved Ash1 FxLP motif as the MRG15 interaction site. (A) Domain architecture of MRG15, Caf1 and Ash1 and alignment of Ash1 protein sequences harbouring the FxLP motif. (B-E) Total extracts (In) from Hi-5 cells co-expressing the indicated AMC subunits and material isolated from these cells (Elu) by Strep- (B-D) or His- (E) affinity purification, separated on 8% (B,C,E) or 10% (D) SDS polyacrylamide gels and visualized by Coomassie Blue staining. (E) Use of the His-tag on Caf1 and a different running buffer accounts for the slower migration behaviour of Caf1 relative to MRG15 in this experiment. See text for a description of the results.