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

Nature Structural Biology volume 10, pages 386393 (2003) | Download Citation

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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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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.

Author information

Author notes

    • Tomoo Shiba
    •  & Masato Kawasaki

    These authors have contributed equally to this work.

    • Hiroyuki Takatsu

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

    • Terukazu Nogi

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

    • Kazuhisa Nakayama

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

Affiliations

  1. Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki 305-0801, 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, Ibaraki 305-0062, Japan.

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

    • Hiroyuki Takatsu
    •  & Kazuhisa Nakayama

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The authors declare no competing financial interests.

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Correspondence to Soichi Wakatsuki.

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https://doi.org/10.1038/nsb920

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