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Fig. 4. Plant AP180{delta}ENTH binding to {alpha}C-adaptin ear constructs. (A) Pull-down experiments with the full-length ear region or the truncated version of At-{alpha}C-Ad and GST as a control together with the recombinant H6-AtAP180{delta}ENTH construct show that binding requires the ear region. (B) Analysis of the consensus binding motifs of At-AP180 for binding to the {alpha}C-ear. Binding experiments with alanine substitutions mutans of either the DPF motif (At-AP180{delta}ENTH-DPF/AAA) or the DPW-motif (At-AP180{delta}ENTH-DPW/AAA) with the full ear construct of At-{alpha}C-adaptin reveal that the DPF-motif is crucial for binding. GST serves as a control. The antibody against histidine recognizes the AP180-constructs in immunoblots of (A) and (B). P, pellet; S, supernatant.





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