Development of a bioassay for detection of Wnt-binding affinities for individual frizzled receptors

Anal Biochem. 2010 Jun 15;401(2):288-94. doi: 10.1016/j.ab.2010.03.009. Epub 2010 Mar 12.

Abstract

Wnts are secreted lipid-modified glycoproteins that carry out various signaling functions during development and in adult tissue. Wnt signaling is mediated by frizzled receptors (Fzds) at the cell surface and can be modulated by the secreted frizzled-related proteins (SFRPs) and other molecular antagonists. Abnormal Wnt signaling has been implicated in several diseases. However, due to the complexity of the Wnt signal and the lack of knowledge pertaining to the binding properties of different Wnt ligands, no therapeutic agents that target this pathway exist. Using a novel enzyme-linked immunosorbent assay (ELISA)-based technique, we were able to determine the first measurements of binding affinity for specific Wnt interactions. This study shows that purified Wnt3a, Wnt7a, and Wnt5a have different binding specificities for Fzds and SFRPs.

Publication types

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

MeSH terms

  • Cell Line, Tumor
  • Endometrial Neoplasms / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Frizzled Receptors / metabolism*
  • Humans
  • MAP Kinase Kinase 4 / metabolism
  • Protein Binding
  • Proto-Oncogene Proteins / metabolism
  • Receptors, G-Protein-Coupled / metabolism
  • Wnt Proteins / metabolism*
  • Wnt3 Protein
  • Wnt3A Protein
  • beta Catenin / metabolism

Substances

  • FZD10 protein, human
  • FZD5 protein, human
  • Frizzled Receptors
  • Proto-Oncogene Proteins
  • Receptors, G-Protein-Coupled
  • SFRP4 protein, human
  • WNT3A protein, human
  • WNT7A protein, human
  • Wnt Proteins
  • Wnt3 Protein
  • Wnt3A Protein
  • beta Catenin
  • MAP Kinase Kinase 4