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Figure 1.
Fig. 1. (A) Schematic illustration of the subunit
organization of the AP-2 adaptor heterotetramer. Regions of the
complex with known protein-binding functions are indicated. For
tyrosine-based internalization signals, X is any amino acid and
Ø represents a bulky hydrophobic residue (1, 3). (B)
Functional protein associations with the GST-  [C]-appendage
fusion protein. Purified GST- [C]
appendage (0-100 µg) or GST (100 µg), immobilized on
25 µl packed glutathione Sepharose, were incubated in  7.5 mg/ml rat
brain cytosol for 60 min at 4°C. The Sepharose beads were
then recovered by centrifugation, and aliquots corresponding to
1/150 of the
supernatant (S) and 1/10 of each pellet (P) were resolved by
SDS/PAGE and either stained with Coomassie blue (Left) or
transferred to nitrocellulose (Right). Portions of the blots
were probed with anti-epsin, anti-eps15, anti-amphiphysin,
anti-AP180, or anti-dynamin antibodies. The position of the
markers (kDa) is indicated on the left and only the relevant
portion of each blot is shown.
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