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The structure of SHH in complex with HHIP reveals a recognition role for the Shh pseudo active site in signaling

Nature Structural & Molecular Biology volume 16, pages 691–697 (2009)Cite this article

Abstract

Hedgehog (Hh) signaling is crucial for many aspects of embryonic development, whereas dysregulation of this pathway is associated with several types of cancer. Hedgehog-interacting protein (Hhip) is a surface receptor antagonist that is equipotent against all three mammalian Hh homologs. The crystal structures of human HHIP alone and bound to Sonic hedgehog (SHH) now reveal that HHIP is comprised of two EGF domains and a six-bladed β-propeller domain. In the complex structure, a critical loop from HHIP binds the pseudo active site groove of SHH and directly coordinates its Zn2+ cation. Notably, sequence comparisons of this SHH binding loop with the Hh receptor Patched (Ptc1) ectodomains and HHIP- and PTC1-peptide binding studies suggest a 'patch for Patched' at the Shh pseudo active site; thus, we propose a role for Hhip as a structural decoy receptor for vertebrate Hh.

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Figure 1: Characterization of the Hhip ECD minimal region required for Hh pathway inhibition.
Figure 2: Structure and function of the HHIP–SHH complex.
Figure 3: Shh binding sites and the presence of an 'L2-like loop' in PTC1.
Figure 4: HHIP and PTC1 competitively bind at the SHH pseudo active site groove.
Figure 5: Proposed model of Hhip regulation of the Shh signaling pathway.

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Acknowledgements

We thank our colleagues H. Tian, T. Tang, F. Rousseau, C. Noguère, A. Esteves and C. Quan for reagents and technical advice; D. Arnott and E. Drake for 5E1 mapping and affinities; A. Spura and D. Dayne at ForteBio for their support; S. Fakra at the Advanced Light Source (ALS) for her assistance with calcium identification; and our collaborators at Curis, Inc. for S12 cells. Stanford Synchrotron Radiation Laboratory (SSRL), ALS and the Berkeley Center for Structural Biology are supported by the US Department of Energy, National Institutes of Health and the National Institute of General Medical Sciences.

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  1. Ivan Bosanac and Henry R Maun: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Structural Biology, Genentech, Inc., South San Francisco, California, USA

    Ivan Bosanac & Sarah G Hymowitz

  2. Department of Protein Engineering, Genentech, Inc., South San Francisco, California, USA

    Henry R Maun, Andreas Lingel, J Fernando Bazan & Robert A Lazarus

  3. Department of Molecular Biology, Genentech, Inc., South San Francisco, California, USA

    Suzie J Scales, Xiaohui Wen & Frederic J de Sauvage

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Contributions

I.B. performed structural biology; H.R.M. expressed and purified proteins and performed biochemical assays; S.J.S. and X.W. performed cell-based assays; A.L. contributed to NMR experiments; J.F.B. carried out computational analysis of the Hhip ECD; I.B., H.R.M., S.J.S., A.L., J.F.B., F.J.d.S., S.G.H. and R.A.L. analyzed data and wrote the manuscript. All authors are employees of Genentech, Inc.

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Correspondence to Frederic J de Sauvage, Sarah G Hymowitz or Robert A Lazarus.

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Bosanac, I., Maun, H., Scales, S. et al. The structure of SHH in complex with HHIP reveals a recognition role for the Shh pseudo active site in signaling. Nat Struct Mol Biol 16, 691–697 (2009). https://doi.org/10.1038/nsmb.1632

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