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The SPRY domain of SSB-2 adopts a novel fold that presents conserved Par-4–binding residues

Nature Structural & Molecular Biology volume 13, pages 7784 (2006) | Download Citation

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Abstract

The four mammalian SPRY domain–containing SOCS box proteins (SSB-1 to SSB-4) are characterized by a C-terminal SOCS box and a central SPRY domain. We have determined the first SPRY-domain structure, as part of SSB-2, by NMR. This domain adopts a novel fold consisting of a β-sandwich structure formed by two four-stranded antiparallel β-sheets with a unique topology. We demonstrate that SSB-1, SSB-2 and SSB-4, but not SSB-3, bind prostate apoptosis response protein-4 (Par-4). Mutational analysis of SSB-2 loop regions identified conserved structural determinants for its interaction with Par-4 and the hepatocyte growth factor receptor, c-Met. Mutations in analogous loop regions of pyrin and midline-1 SPRY domains have been shown to cause Mediterranean fever and Opitz syndrome, respectively. Our findings provide a template for SPRY-domain structure and an insight into the mechanism of SPRY-protein interaction.

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Acknowledgements

This work was supported by the National Health and Medical Research Council, Australia (Program grant 257500), and by AMRAD operations Pty. Ltd., Melbourne, Australia. S.E.N. was supported by a National Health and Medical Research Council Biomedical Career Development award. The authors would like to thank N. Sprigg for expert technical assistance, R. Simpson, L. Connelly and D. Frecklington for protein identification by peptide mass spectroscopy, R. Johnstone for generously providing Par-4 expression constructs and D. Keizer for advice on structure calculations. We also thank P. Colman and W. Alexander for reviewing this manuscript.

Author information

Author notes

    • Seth L Masters
    •  & Shenggen Yao

    These authors contributed equally to this work.

Affiliations

  1. The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, 3050, Victoria, Australia.

    • Seth L Masters
    • , Shenggen Yao
    • , Tracy A Willson
    • , Jian-Guo Zhang
    • , Kirsten R Palmer
    • , Brian J Smith
    • , Jeffrey J Babon
    • , Nicos A Nicola
    • , Raymond S Norton
    •  & Sandra E Nicholson
  2. The Department of Medical Biology, The University of Melbourne, Parkville, 3010, Victoria, Australia.

    • Seth L Masters

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Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Raymond S Norton or Sandra E Nicholson.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    1H-15N HSQC spectrum of SSB-2.

  2. 2.

    Supplementary Fig. 2

    The solution structure of SSB-2.

  3. 3.

    Supplementary Table 1

    Mutational analysis of the SSB-2 SPRY domain

  4. 4.

    Supplementary Table 2

    Primers used for cDNA cloning

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DOI

https://doi.org/10.1038/nsmb1034

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