FIGURE 5.

Clavesins form a complex with AP-1 and CHC. A, GST fusion proteins encoding clavesin 1 or 2 C-terminal regions or GST alone were precoupled to glutathione-Sepharose and incubated with soluble brain extracts. Affinity-selected proteins were blotted with the indicated antibodies. The starting material (SM) is an aliquot of extract equal to one-tenth of that added to the fusion proteins. B, as in A except using GST-full-length clavesin 1 and 2 and full-length constructs lacking the clathrin box (ΔCB). C, a GST fusion protein encoding the N-terminal domain of the CHC (GST-CHC-TD) or GST alone was precoupled to glutathione-Sepharose and incubated with soluble extracts from HEK-293 cells transfected with FLAG-tagged clavesin 1 or 2. Affinity-selected proteins were blotted with an anti-FLAG antibody.