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A Novel Subtype of AP-1-binding Motif within the Palmitoylated trans-Golgi Network/Endosomal Accessory Protein Gadkin/γ-BAR

T. Maritzen, M.R. Schmidt, V. Kukhtina, V.A. Higman, H. Strauss, R. Volkmer, H. Oschkinat, C.G. Dotti, V. Haucke – 2010

Membrane traffic between the trans-Golgi network (TGN) and endosomes is mediated in part by the assembly of clathrin-AP-1 adaptor complex-coated vesicles. This process involves multiple accessory proteins that directly bind to the ear domain of AP-1γ via degenerate peptide motifs that conform to the consensus sequence ØG(P/D/E)(Ø/L/M) (with Ø being a large hydrophobic amino acid). Recently, γ-BAR (hereafter referred to as Gadkin for reasons explained below) has been identified as a novel AP-1 recruitment factor involved in AP-1-dependent endosomal trafficking of lysosomal enzymes. How precisely Gadkin interacts with membranes and with AP-1γ has remained unclear. Here we show that Gadkin is an S-palmitoylated peripheral membrane protein that lacks stable tertiary structure. S-Palmitoylation is required for the recruitment of Gadkin to TGN/endosomal membranes but not for binding to AP-1. Furthermore, we identify a novel subtype of AP-1-binding motif within Gadkin that specifically associates with the γ1-adaptin ear domain. Mutational inactivation of this novel type of motif, either alone or in combination with three more conventional AP-1γ binding peptides, causes Gadkin to mislocalize to the plasma membrane and interferes with its ability to render AP-1 brefeldin A-resistant, indicating its physiological importance. Our studies thus unravel the molecular basis for Gadkin-mediated AP-1 recruitment to TGN/endosomal membranes and identify a novel subtype of the AP-1-binding motif.

Titel
A Novel Subtype of AP-1-binding Motif within the Palmitoylated trans-Golgi Network/Endosomal Accessory Protein Gadkin/γ-BAR
Verfasser
T. Maritzen, M.R. Schmidt, V. Kukhtina, V.A. Higman, H. Strauss, R. Volkmer, H. Oschkinat, C.G. Dotti, V. Haucke
Datum
2010
Kennung
10.1074/jbc.M109.049197
Zitierweise
J. Biol. Chem. 2010, 285(6), 4074-4086
Art
Text
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