A clathrin-binding site in the hinge of the beta 2 chain of mammalian AP-2 complexes - PubMed (original) (raw)
. 1995 Dec 29;270(52):31083-90.
doi: 10.1074/jbc.270.52.31083.
Affiliations
- PMID: 8537368
- DOI: 10.1074/jbc.270.52.31083
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A clathrin-binding site in the hinge of the beta 2 chain of mammalian AP-2 complexes
W Shih et al. J Biol Chem. 1995.
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Abstract
The assembly of cytosolic clathrin into the cytoplasmic face of coated pits and coated vesicles appears to be driven by the clathrin-associated protein (AP) complexes. We have previously shown that one of the large chains of the AP complexes, the beta chain, is sufficient to drive coat assembly in vitro. This chain consists of two domains, the amino-terminal trunk and the carboxyl-terminal ear, linked by a "hinge." We report here that presence of the hinge in recombinant beta trunk or in recombinant beta ear fragments is essential for driving in vitro assembly of clathrin into coats. We have also used a binding assay to map the clathrin-binding site by nested deletion of hinge sequences to a 50-residue region in the center of the hinge. This sequence is conserved in all known beta sequences from multicellular organisms. The interaction of a single beta hinge with a clathrin triskelion is weak, and we propose that recruitment of cytosolic clathrin to a forming coated pit involves simultaneous contacts between the legs of single clathrin trimers and the beta hinges of two or three membrane-bound AP complexes. Uncoating is likely to require interruption of these contacts.
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