GGA1 interacts with the adaptor protein AP-1 through a WNSF sequence in its hinge region

J Biol Chem. 2004 Apr 23;279(17):17411-7. doi: 10.1074/jbc.M401158200. Epub 2004 Feb 18.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ADP-Ribosylation Factors / chemistry
  • ADP-Ribosylation Factors / metabolism*
  • Adaptor Proteins, Vesicular Transport*
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Binding, Competitive
  • Brain / metabolism
  • COS Cells
  • Carrier Proteins / chemistry
  • Carrier Proteins / metabolism*
  • DNA, Complementary / metabolism
  • Dose-Response Relationship, Drug
  • Escherichia coli / metabolism
  • Gene Library
  • Glutathione Transferase / metabolism
  • Humans
  • Immunoblotting
  • In Vitro Techniques
  • Membrane Proteins / chemistry
  • Molecular Sequence Data
  • Mutation
  • Nerve Tissue Proteins / chemistry
  • Peptides / chemistry
  • Phenylalanine / chemistry
  • Plasmids / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • Receptor, IGF Type 2 / chemistry
  • Sequence Homology, Amino Acid
  • Transcription Factor AP-1 / metabolism*
  • Tryptophan / chemistry
  • Vesicular Transport Proteins*
  • trans-Golgi Network / metabolism

Substances

  • Adaptor Proteins, Vesicular Transport
  • Carrier Proteins
  • DNA, Complementary
  • GGA adaptor proteins
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Peptides
  • RABEP2 protein, human
  • Receptor, IGF Type 2
  • Transcription Factor AP-1
  • Vesicular Transport Proteins
  • amphiphysin
  • Phenylalanine
  • Tryptophan
  • Glutathione Transferase
  • ADP-Ribosylation Factors