Journal article
Structural basis for Par-4 recognition by the SPRY domain- and SOCS box-containing proteins SPSB1, SPSB2, and SPSB4
P Filippakopoulos, A Low, TD Sharpe, J Uppenberg, S Yao, Z Kuang, P Savitsky, RS Lewis, SE Nicholson, RS Norton, AN Bullock
Journal of Molecular Biology | ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD | Published : 2010
Open access
Abstract
The mammalian SPRY domain- and SOCS box-containing proteins, SPSB1 to SPSB4, belong to the SOCS box family of E3 ubiquitin ligases. Substrate recognition sites for the SPRY domain are identified only for human Par-4 (ELNNNL) and for the Drosophila orthologue GUSTAVUS binding to the DEAD-box RNA helicase VASA (DINNNN). To further investigate this consensus motif, we determined the crystal structures of SPSB1, SPSB2, and SPSB4, as well as their binding modes and affinities for both Par-4 and VASA. Mutation of each of the three Asn residues in Par-4 abrogated binding to all three SPSB proteins, while changing EL to DI enhanced binding. By comparison to SPSB1 and SPSB4, the more divergent protei..
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Awarded by GlaxoSmithKline
Funding Acknowledgements
This work was supported in part by the National Health and Medical Research Council, Australia (Program grant 461219 and Project grant 461233 to R.S.N. and S.E.N., fellowships to R.S.N. and S.E.N., and a National Health and Medical Research Council IRIISS grant 361646), as well as a Victorian State Government OIS grant. We thank Brian Smith for his assistance with the molecular dynamics simulations. We thank Frank von Delft as well as staff at the Swiss Light Source and Diamond synchrotrons for assistance with diffraction data collection. The Structural Genomics Consortium is a registered charity (number 1097737) that receives funds from the Canadian Institutes for Health Research, the Canadian Foundation for Innovation, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Karolinska Institutet, the Knut and Alice Wallenberg Foundation, the Ontario Innovation Trust, the Ontario Ministry for Research and Innovation, Merck & Co Inc., the Novartis Research Foundation, the Swedish Agency for Innovation Systems, the Swedish Foundation for Strategic Research, and the Wellcome Trust.