The ∼16 kDa C-Terminal Sequence of Clathrin Assembly Protein AP180 Is Essential for Efficient Clathrin Binding
Figure 3
The ∼16 kDa C-terminal sub-domain is required for clathrin binding.
(A) C½, C½-T1, C½-T2, C½-T3, C½-T4 and C½-T5 were used in pull-downs with rat brain lysate (separate rats for each duplicate lane). Each sample was run on a large format 10% SDS-PAGE gel, transferred onto nitrocellulose membrane and probed with a CHC antibody. (B) One eighth of the amount used to produce (A) was loaded onto a mini-gel and stained with Coomassie blue to show the relative amount of GST fusion protein used as bait. (C) AP180 FL, FL-T1, FL-T2, FL-T3, FL-T4 and FL-T5 were used in pull-down experiments with rat brain lysate and probed with a CHC antibody. (D) One eighth of the amount used to produce (C) was loaded onto a mini-gel and stained with Coomassie blue to show the relative amount of GST fusion protein used as bait.