Fig. 3. Features of AP180 homologs and At-AP180 constructs. (A) Full-length At-AP180 comprises an N-terminal ENTH-domain and two putative binding sites for the -adaptin ear region (⁴⁷³DPW, ⁶⁰⁸DPF), as well as one clathrin-binding consensus motif (⁴⁴²DLL). The histidine or GST-fusion protein constructs of At-AP180, lacking the ENTH domain (H₆/GST-At-AP180ENTH), were used in pull-down and in clathrin assembly experiments. (B) Commassie-stained gel of the purification of the histidine-tagged recombinant full-length At-AP180 protein (predicted Mr is 72x10³; lane 1, marked by a star) and also consecutive stepwise purification of the H₆-At-AP180ENTH fusion-protein (predicted Mr is 34.5x10³) after incubation with Ni-NTA-beads (2) then boiled (3) and then after gelfiltration on Superose 6/FPLC (4). The purified GST-AP180ENTH fusion-protein has a molecular mass of 55 kDa (5).