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Recognition of accessory protein motifs by the γ-adaptin ear domain of GGA3

Nature Structural & Molecular Biology volume 10, pages 599–606 (2003)Cite this article

Abstract

Adaptor proteins load transmembrane protein cargo into transport vesicles and serve as nexuses for the formation of large multiprotein complexes on the nascent vesicles. The γ-adaptin ear (GAE) domains of the AP-1 adaptor protein complex and the GGA adaptor proteins recruit accessory proteins to these multiprotein complexes by binding to a hydrophobic motif. We determined the structure of the GAE domain of human GGA3 in complex with a peptide based on the DFGPLV sequence of the accessory protein Rabaptin-5 and refined it at a resolution of 2.2 Å. The leucine and valine residues of the peptide are partly buried in two contiguous shallow, hydrophobic depressions. The anchoring phenylalanine is buried in a deep pocket formed by the aliphatic portions of two conserved arginine residues, along with an alanine and a proline, illustrating the unusual function of a cluster of basic residues in binding a hydrophobic motif.

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Figure 1: Binding of Rabaptin-5, γ-synergin and enthoprotin peptides to GGA3(571–723).
Figure 2: Binding of GGA3-GAE mutants to Rabaptin-5.
Figure 3: Structure determination of the GAE domain–peptide complex.
Figure 4: Overall structure of the GGA3-GAE domains from GGA3 and comparison to the γ1-adaptin ear.
Figure 5
Figure 6: Conservation of GAE domains.

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Protein Data Bank

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Acknowledgements

We thank T. Earnest and the staff of the HHMI beamline 8.2.1, Advanced Light Source, Lawrence Berkeley Lab (Berkeley, California) for assistance with data collection, S. Sechi for assistance with mass spectrometry, and X. Zhu for expert technical assistance.

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Author notes

  1. Gregory J Miller and Rafael Mattera: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Health and Human Services, Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Gregory J Miller & James H Hurley

  2. Department of Health and Human Services, Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Rafael Mattera & Juan S Bonifacino

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  1. Gregory J Miller
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Correspondence to James H Hurley.

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Miller, G., Mattera, R., Bonifacino, J. et al. Recognition of accessory protein motifs by the γ-adaptin ear domain of GGA3. Nat Struct Mol Biol 10, 599–606 (2003). https://doi.org/10.1038/nsb953

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