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Molecular mechanism of membrane recruitment of GGA by ARF in lysosomal protein transport

Nature Structural & Molecular Biology volume 10, pages 386–393 (2003)Cite this article

Abstract

GGAs are critical for trafficking soluble proteins from the trans-Golgi network (TGN) to endosomes/lysosomes through interactions with TGN-sorting receptors, ADP-ribosylation factor (ARF) and clathrin. ARF–GTP bound to TGN membranes recruits its effector GGA by binding to the GAT domain, thus facilitating recognition of GGA for cargo-loaded receptors. Here we report the X-ray crystal structures of the human GGA1-GAT domain and the complex between ARF1–GTP and the N-terminal region of the GAT domain. When unbound, the GAT domain forms an elongated bundle of three a-helices with a hydrophobic core. Structurally, this domain, combined with the preceding VHS domain, resembles CALM, an AP180 homolog involved in endocytosis. In the complex with ARF1–GTP, a helix-loop-helix of the N-terminal part of GGA1-GAT interacts with the switches 1 and 2 of ARF1 predominantly in a hydrophobic manner. These data reveal a molecular mechanism underlying membrane recruitment of adaptor proteins by ARF–GTP.

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Figure 1: Structures of the GGA1-GAT domain and its complex with ARF1.
Figure 2: Comparison of the GGA VHS-GAT domain with AP180 homologs.
Figure 3: SPR and CD measurements of N-GAT and GAT domains.
Figure 4: Interaction between ARF1 and GGA1 N-GAT.
Figure 5: Comparison of the ARF1–N-GAT complex with other complex structures of ARF and ARF-interacting proteins shown in stereo diagrams: a, the ARF1–N-GAT complex (this work); b, ARF1–ARFGAP complex20; c, ARF1–Sec7 domain complex21; and d, Arl2–PDEδ complex23.
Figure 6: Domain organization of GGA and a proposed model of the interactions with its partners at several stages of the vesicle formation: M6PR, ARF–GTP, clathrin N-terminal propeller and an accessory protein.

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Acknowledgements

Coordinates of the two complex structures, ARF–ARFGAP and ARF–Sec7 domain, were provided by J. Goldberg. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, from the Japan Society for Promotion of Science (fellowship to H.T.), from the University of Tsukuba Research Projects, and by Protein 3000 Project of the MEXT.

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Author notes

  1. Hiroyuki Takatsu

    Present address: RIKEN Research Center for Allergy and Immunology (RCAI), Yokohama, Kanagawa, 230-0045, Japan

  2. Terukazu Nogi

    Present address: Max-Planck-Institute of Biophysics, 60439, Frankfurt am Main, Germany

  3. Kazuhisa Nakayama

    Present address: Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Ishikawa, 920-0934, Japan

  4. Tomoo Shiba, Masato Kawasaki, Hiroyuki Takatsu and Kazuhisa Nakayama: These authors have contributed equally to this work.

Authors and Affiliations

  1. Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK), Tsukuba, 305-0801, Ibaraki, Japan

    Tomoo Shiba, Masato Kawasaki, Terukazu Nogi, Naohiro Matsugaki, Noriyuki Igarashi, Mamoru Suzuki, Ryuichi Kato & Soichi Wakatsuki

  2. Foundation for Advancement of International Science (FAIS), Tsukuba, 305-0062, Ibaraki, Japan

    Tomoo Shiba

  3. Institute of Biological Sciences and Gene Research Center, University of Tsukuba, Tsukuba, 305-8572, Ibaraki, Japan

    Hiroyuki Takatsu & Kazuhisa Nakayama

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Shiba, T., Kawasaki, M., Takatsu, H. et al. Molecular mechanism of membrane recruitment of GGA by ARF in lysosomal protein transport. Nat Struct Mol Biol 10, 386–393 (2003). https://doi.org/10.1038/nsb920

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