Identification of the clathrin-binding domain of assembly protein AP-2.
The clathrin binding domain of the assembly protein AP-2 has been identified by proteolytically cleaving AP-2 into 2 discrete moieties, termed light and heavy mero-AP (LM-AP and HM-AP), and testing their ability to bind to clathrin assembled into cage structures or to clathrin trimers immobilized on Sepharose. The smaller product (LM-AP), which contains 20-40-kD fragments of the parent 100-kD polypeptides and which comprises two small appendages in the native AP-2 molecule, did not significantly interact with clathrin under either condition. In contrast, the HM-AP complex, which forms the larger central mass of the native AP-2 structure and contains uncleaved 50-kD and 16-kD polypeptides as well as 60-66-kD fragments of the parent 100-kD polypeptides, retained binding activity for both dissociated and assembled clathrin.
Keen
J H
JH
Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140.
Beck
K A
KA
D033942:Adaptor Proteins, Vesicular Transport; D001665:Binding Sites; D002846:Chromatography, Affinity; D002966:Clathrin / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding
D033942:Adaptor Proteins, Vesicular Transport; D002966:Clathrin; D010750:Phosphoproteins