GGA1 interacts with the adaptor protein AP-1 through a WNSF sequence in its hinge region.
The Golgi-associated gamma-adaptin-related ADP-ribosylation factor-binding proteins (GGAs) are critical components of the transport machinery that mediates the trafficking of the mannose 6-phosphate receptors and associated cargo from the trans-Golgi network to the endosomes. The GGAs colocalize in vivo with the clathrin adaptor protein AP-1 and bind to AP-1 in vitro, suggesting that the two proteins may cooperate in packaging the mannose 6-phosphate receptors into clathrin-coated vesicles at the trans-Golgi network. Here, we demonstrate that the sequence, (382)WNSF(385), in the hinge region of GGA1 mediates its interaction with the AP-1 gamma-ear. The Trp and Phe constitute critical amino acids in this interaction. The binding of Rabaptin5 to the AP-1 gamma-ear, which occurs through a FXXPhi motif, is inhibited by a peptide encoding the GGA1 (382)WNSF(385) sequence. Moreover, mutations in the AP-1 gamma-ear that abolish its interaction with Rabaptin5 also preclude its association with GGA1. These results suggest that the GGA1 WXXF-type and Rabaptin5 FXXPhi-type motifs bind to the same or highly overlapping sites in the AP-1 gamma-ear. This binding is modulated by residues adjacent to the core motifs.
Pubmed ID: 14973137 RIS Download
ADP-Ribosylation Factors | Adaptor Proteins, Vesicular Transport | Amino Acid Motifs | Amino Acid Sequence | Animals | Binding Sites | Binding, Competitive | Brain | COS Cells | Carrier Proteins | DNA, Complementary | Dose-Response Relationship, Drug | Escherichia coli | Gene Library | Glutathione Transferase | Humans | Immunoblotting | In Vitro Techniques | Membrane Proteins | Molecular Sequence Data | Mutation | Nerve Tissue Proteins | Peptides | Phenylalanine | Plasmids | Protein Binding | Protein Structure, Tertiary | Receptor, IGF Type 2 | Sequence Homology, Amino Acid | Transcription Factor AP-1 | Tryptophan | Vesicular Transport Proteins | trans-Golgi Network