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.
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Supplementary information
Supplementary Fig. 1
1H-15N HSQC spectrum of SSB-2. (PDF 141 kb)
Supplementary Fig. 2
The solution structure of SSB-2. (PDF 107 kb)
Supplementary Table 1
Mutational analysis of the SSB-2 SPRY domain (PDF 35 kb)
Supplementary Table 2
Primers used for cDNA cloning (PDF 1194 kb)
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Masters, S., Yao, S., Willson, T. et al. The SPRY domain of SSB-2 adopts a novel fold that presents conserved Par-4–binding residues. Nat Struct Mol Biol 13, 77–84 (2006). https://doi.org/10.1038/nsmb1034
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DOI: https://doi.org/10.1038/nsmb1034
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