Structure of Protein Related to Dan and Cerberus: Insights Into the Mechanism of Bone Morphogenetic Protein Antagonism

Structure. 2013 Aug 6;21(8):1417-29. doi: 10.1016/j.str.2013.06.005. Epub 2013 Jul 11.

Abstract

The bone morphogenetic proteins (BMPs) are secreted ligands largely known for their functional roles in embryogenesis and tissue development. A number of structurally diverse extracellular antagonists inhibit BMP ligands to regulate signaling. The differential screening-selected gene aberrative in neuroblastoma (DAN) family of antagonists represents the largest group of BMP inhibitors; however, little is known of how they mechanistically inhibit BMP ligands. Here, we present the structure of the DAN family member, protein related to Dan and Cerberus (PRDC), solved by X-ray crystallography. The structure reveals a growth factor-like appearance with an unexpected dimerization mechanism that is formed through extensive β strand contacts. Using site-directed mutagenesis coupled with in vitro and in vivo activity assays, we identified a BMP-binding epitope on PRDC. We also determined that PRDC binds heparin with high affinity and that heparin binding to PRDC interferes with BMP antagonism. These results offer insight for how DAN family antagonists functionally inhibit BMP ligands.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • Bone Morphogenetic Protein 2 / antagonists & inhibitors
  • Bone Morphogenetic Protein 2 / chemistry*
  • Bone Morphogenetic Protein 2 / physiology
  • Cell Cycle Proteins
  • Crystallography, X-Ray
  • Cytokines
  • Heparin / chemistry
  • Intercellular Signaling Peptides and Proteins / chemistry
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Multimerization
  • Protein Structure, Secondary
  • Proteins / chemistry*
  • Proteins / genetics
  • Proteins / physiology
  • Scattering, Small Angle
  • Structural Homology, Protein
  • Surface Properties
  • Xenopus Proteins / chemistry
  • Xenopus laevis

Substances

  • Bmp2 protein, mouse
  • Bone Morphogenetic Protein 2
  • Cell Cycle Proteins
  • Cytokines
  • Grem2 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • Nbl1 protein, mouse
  • Proteins
  • Xenopus Proteins
  • cer1 protein, Xenopus
  • Heparin

Associated data

  • PDB/4JPH