Proteomic Analysis of Adaptor Protein 1A Coats Selectively Assembled on Liposomes

Coat components localize to specific membrane domains, where they sort selected transmembrane proteins. To study how clathrin coats are stabilized on such domains and to identify the protein networks involved, we combined proteomic screens and in vitro liposome-based assays that recapitulate the fid...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2006-02, Vol.103 (9), p.3159-3164
Main Authors: Baust, Thorsten, Czupalla, Cornelia, Krause, Eberhard, Bourel-Bonnet, Line, Hoflack, Bernard
Format: Article
Language:English
Subjects:
Actins
Actins - metabolism
Adaptor Proteins, Signal Transducing - chemistry
Adaptor Proteins, Signal Transducing - genetics
Adaptor Proteins, Signal Transducing - metabolism
Amino Acid Sequence
Animals
Binding sites
Biological Sciences
Cell Membrane - metabolism
Cell membranes
Cells
Cellular biology
Cytosol
Endosomes
Liposomes
Liposomes - chemistry
Liposomes - metabolism
Membrane Fusion
Membranes
Molecular Sequence Data
P branes
Phosphatidylinositol Phosphates - pharmacology
Polymerization
Protein Binding
Proteins
Proteomics
rab GTP-Binding Proteins - metabolism
Racism
Swine
Transmembrane proteins
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Summary:Coat components localize to specific membrane domains, where they sort selected transmembrane proteins. To study how clathrin coats are stabilized on such domains and to identify the protein networks involved, we combined proteomic screens and in vitro liposome-based assays that recapitulate the fidelity of protein sorting in vivo. Our study identifying ≈40 proteins on AP-1Acoated liposomes revealed that AP-1A coat assembly triggers the concomitant recruitment of Raci, its effectors, and the Wave/Scar complex as well as that of Rab11 and Rabl4. The coordinated recruitment of these different machineries requires a mosaic of membrane components comprising the GTPase ADP-ribosylation factor 1, sorting signals in selected transmembrane proteins, and phosphatidylinositol 4-phosphate. These results demonstrate that the combinatorial use of low-affinity binding sites present on the same membrane domain accounts not only for a selective coat assembly but also for the coordinated assembly of selected machineries required for actin polymerization and subsequent membrane fusion.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0511062103