Cooperative folding and ligand-binding properties of LRP6 beta-propeller domains

J Biol Chem. 2009 May 29;284(22):15299-307. doi: 10.1074/jbc.M807285200. Epub 2009 Mar 31.

Abstract

Wnt/beta-catenin signaling controls cell growth during development, and its misregulation in adults can cause human diseases. LRP6, the essential co-receptor for the Wnt pathway, consists of four beta-propeller domains flanked by epidermal growth factor repeats in its extracellular region. To understand the maturation and ligand-binding properties of individual BP domains, we generated soluble receptor consisting of individual BPs, as well as combinations of these domains. We show that BP1, BP2, and BP4 each can be folded and secreted, and their secretion was enhanced by co-expression of Mesd, a molecular chaperone essential for LRP6 folding and maturation. BP3 is not secreted when expressed on its own or in combination with BP2 or BP1 and 2 (BP12); however, folding and secretion of BP3 is vastly enhanced when expressed together with BP4. Similar cooperative folding and maturation was observed between BP1 and BP2. These results suggest that BP1 forms a functional folding unit with BP2, whereas BP3 folds together with BP4. Using these BP constructs, we also found that BP12 and BP34 constitute independent ligand-binding domains capable of binding Wnt3a, Dkk1, and Mesd. The ability of Mesd to block the binding of both Wnt3a and Dkk1 to LRP6 enables this specialized chaperone to function as a Wnt signaling modulator. Together, our studies reveal unique properties of the LRP6 BP domains and provide novel tools to understand LRP6 function in ligand binding and Wnt signaling. Our results also support the development of soluble LRP6 receptors and Mesd as potential therapeutic molecules that target Wnt signaling.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Culture Media, Conditioned
  • Epidermal Growth Factor / metabolism
  • Gene Knockdown Techniques
  • Humans
  • LDL-Receptor Related Proteins / chemistry*
  • LDL-Receptor Related Proteins / metabolism*
  • Ligands
  • Low Density Lipoprotein Receptor-Related Protein-6
  • Mice
  • Molecular Chaperones / metabolism
  • Protein Binding
  • Protein Folding*
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Signal Transduction
  • Wnt Proteins / metabolism

Substances

  • Culture Media, Conditioned
  • LDL-Receptor Related Proteins
  • LRP6 protein, human
  • Ligands
  • Low Density Lipoprotein Receptor-Related Protein-6
  • Lrp6 protein, mouse
  • Molecular Chaperones
  • Wnt Proteins
  • Epidermal Growth Factor