Expression and Purification of Recombinant Protein Related to DAN and Cerberus (PRDC)

Protein Expr Purif. 2012 Apr;82(2):389-95. doi: 10.1016/j.pep.2012.02.010. Epub 2012 Feb 20.

Abstract

Bone morphogenetic proteins (BMPs) are secreted protein ligands that control numerous biological processes, such as cell differentiation and cell proliferation. Ligands are regulated by a large number of structurally diverse extracellular antagonists. PRDC or protein related to DAN and cerberus is a BMP antagonist of the DAN family, which is defined by a conserved pattern of cysteine residues that form a ring structure. Here we present the expression and purification of recombinant mouse PRDC (mPRDC) from bacterial (Escherichia coli) inclusion bodies through oxidative refolding. Functional mPRDC was isolated from a nonfunctional component through reverse phase chromatography and shown to inhibit BMP2 and BMP4 in a cell-based luciferase reporter assay. Recombinant mPRDC also bound directly to BMP2, BMP4 and BMP7, but not activin A. Furthermore, circular dichroism indicated that mPRDC is folded and contains a higher than anticipated helical content for a DAN family member protein.

Publication types

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

MeSH terms

  • Animals
  • Bone Morphogenetic Proteins / antagonists & inhibitors
  • Bone Morphogenetic Proteins / physiology
  • Cell Line
  • Chromatography, Reverse-Phase
  • Cytokines
  • Gene Expression
  • Genes, Reporter
  • Hydrophobic and Hydrophilic Interactions
  • Inclusion Bodies / chemistry
  • Luciferases / biosynthesis
  • Luciferases / genetics
  • Mice
  • Protein Binding
  • Protein Refolding
  • Protein Structure, Secondary
  • Proteins / chemistry
  • Proteins / isolation & purification
  • Proteins / metabolism*
  • Proteins / pharmacology
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Recombinant Proteins / pharmacology
  • Signal Transduction
  • Transforming Growth Factor beta / chemistry

Substances

  • Bone Morphogenetic Proteins
  • Cytokines
  • Grem2 protein, mouse
  • Proteins
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • Luciferases