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Interaction of amphiphysins with AP-1 clathrin adaptors at the membrane

Huser, Sonja and Suri, Gregor and Crottet, Pascal and Spiess, Martin. (2013) Interaction of amphiphysins with AP-1 clathrin adaptors at the membrane. Biochemical journal, Vol. 450, Pt. 1. pp. 73-83.

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Official URL: http://edoc.unibas.ch/dok/A6070441

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Abstract

The assembly of clathrin/AP-1-coated vesicles on the trans-Golgi network and endosomes is much less studied than of clathrin/AP-2 vesicles at the plasma membrane for endocytosis. In vitro, AP-1 association to protein-free liposomes had been shown to require phosphoinositides, Arf1•GTP, and additional cytosolic factor(s). We have purified an active fraction from brain cytosol and found it to contain amphiphysin 1 and 2 and endophilin A1, three proteins known to be involved in the formation of AP-2/clathrin coats at the plasma membrane. Assays with bacterially expressed and purified proteins showed AP-1 stabilization on liposomes to depend on amphiphysin 2 or the amphiphysin 1/2 heterodimer. Activity is independent of the SH3 domain, but requires the WDLW motif interacting with γ-adaptin. Endogenous amphiphysin in neurons and transfected protein in cell lines colocalize perinuclearly with AP-1 at the trans-Golgi network. This localization depends on the clathrin and adaptor interaction sequence in the amphiphysins and is sensitive to brefeldin A, which inhibits Arf1-dependent AP-1 recruitment. Interaction between AP-1 and amphiphysin 1/2 in vivo was demonstrated by coimmunoprecipitation after crosslinking. These results suggest an involvement of amphiphysins not only with AP-2 at the plasma membrane, but also in AP-1/clathrin coat formation at the trans-Golgi network.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Growth & Development > Biochemistry (Spiess)
UniBasel Contributors:Spiess, Martin
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:Portland Press
ISSN:0264-6021
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:24 May 2013 09:21
Deposited On:24 May 2013 08:59

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