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Structural basis for binding of accessory proteins by the appendage domain of GGAs

Nature Structural & Molecular Biology volume 10, pages 607–613 (2003)Cite this article

Abstract

The Golgi-associated, γ-adaptin-related, ADP-ribosylation-factor binding proteins (GGAs) and adaptor protein (AP)-1 are adaptors involved in clathrin-mediated transport between the trans-Golgi network and endosomal system. The appendage domains of GGAs and the AP-1 γ-adaptin subunit are structurally homologous and have been proposed to bind to accessory proteins via interaction with short sequences containing phenylalanines and acidic residues. Here we present the structure of the human GGA1 appendage in complex with its cognate binding peptide from the p56 accessory protein (DDDDFGGFEAAETFD) as determined by X-ray crystallography. The interaction is governed predominantly by packing of the first two phenylalanine residues of the peptide with conserved basic and hydrophobic residues from GGA1. Additionally, several main chain hydrogen bonds cause the peptide to form an additional β-strand on the edge of the preexisting β-sheet of the protein. Isothermal titration calorimetry was used to assess the affinities of different peptides for the GGA and γ-appendage domains.

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Figure 1: Schematic models of adaptor proteins and sequences of appendage domain binding motifs.
Figure 2: Structure of the GGA1 appendage in complex with the DDDDFGGFEAAETFD binding peptide from p56.
Figure 3: Details of the GGA1 appendage–p56 peptide interaction.
Figure 4: ITC binding studies of accessory protein peptides to the GGA and γ-appendage domains.

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Acknowledgements

We thank H. Kent for the γ-appendage DNA, J. Hurley and associates for sharing information prior to publication and M. Noble for helpful discussions. G.J.K.P. is funded by a Marie Curie Fellowship of the European Union. D.J.O. is funded by a Wellcome Trust Senior Research Fellowship in Basic Biomedical Science. M.S.R. is funded by a Wellcome Trust Principal Research Fellowship.

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  1. Department of Clinical Biochemistry, Cambridge Institute for Medical Research, University of Cambridge, Hills Road, Cambridge, CB2 2XY, UK

    Brett M Collins, Margaret S Robinson & David J Owen

  2. Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK

    Gerrit J K Praefcke

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Correspondence to Brett M Collins.

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Collins, B., Praefcke, G., Robinson, M. et al. Structural basis for binding of accessory proteins by the appendage domain of GGAs. Nat Struct Mol Biol 10, 607–613 (2003). https://doi.org/10.1038/nsb955

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