Detergent-solubilized Patched purified from Sf9 cells fails to interact strongly with cognate Hedgehog or Ihog homologs

Protein Expr Purif. 2014 Dec;104:92-102. doi: 10.1016/j.pep.2014.09.012. Epub 2014 Sep 26.


Patched (Ptc) is a twelve-pass transmembrane protein that functions as a receptor for the Hedgehog (Hh) family of morphogens. In addition to Ptc, several accessory proteins including the CDO/Ihog family of co-receptors are necessary for proper Hh signaling. Structures of Hh proteins bound to members of the CDO/Ihog family are known, but the nature of the full Hh receptor complex is not well understood. We have expressed the Drosophila Patched and Mouse Patched-1 proteins in Sf9 cells and find that Sonic Hedgehog will bind to Mouse Patched-1 in isolated Sf9 cell membranes but that purified, detergent-solubilized Ptc proteins do not interact strongly with cognate Hh and CDO/Ihog homologs. These results may reflect a nonnative conformation of detergent-solubilized Ptc or that an additional factor or factors lost during purification are required for high-affinity Ptc binding to Hh.

Keywords: Hedgehog signaling; Patched; Protein purification.

Publication types

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

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Detergents / chemistry*
  • Drosophila Proteins / chemistry*
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / isolation & purification
  • Hedgehog Proteins / chemistry*
  • Membrane Glycoproteins / chemistry*
  • Mice
  • Patched-1 Receptor / chemistry
  • Patched-1 Receptor / genetics*
  • Patched-1 Receptor / isolation & purification
  • Protein Binding
  • Protein Conformation
  • Receptors, Cell Surface / chemistry*
  • Receptors, Cell Surface / genetics*
  • Receptors, Cell Surface / isolation & purification
  • Sf9 Cells
  • Solubility


  • Detergents
  • Drosophila Proteins
  • Hedgehog Proteins
  • Membrane Glycoproteins
  • Patched-1 Receptor
  • Receptors, Cell Surface
  • iHog protein, Drosophila
  • ptc protein, Drosophila