Solubilization and functional reconstitution of human neuropeptide FF2 receptors

Anal Biochem. 2010 Mar 15;398(2):225-9. doi: 10.1016/j.ab.2009.12.012. Epub 2009 Dec 14.

Abstract

Neuropeptide FF (NPFF, FLFQPQRFamide) receptors modulate endogenous opioid functions. Here, we report the solubilization of the human NPFF(2) receptor expressed in Chinese hamster ovary (CHO) cells by the zwitterionic detergent Chaps. Chaps solubilization resulted in the abolishment of specific agonist binding activity, which was restored by a polyethylene glycol (PEG) precipitation method. Reincorporation after the precipitation step into liposomes made of endogenous lipids issued from CHO membranes or exogenous lipids significantly enhanced the specific agonist binding activity and G-protein coupling. This method of solubilization and lipid reconstitution could be useful for studies of NPFF receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cholic Acids / pharmacology
  • Cricetinae
  • Cricetulus
  • Detergents / pharmacology
  • Gene Expression / drug effects
  • Humans
  • Receptors, Neuropeptide / chemistry*
  • Receptors, Neuropeptide / genetics
  • Receptors, Neuropeptide / metabolism*
  • Solubility

Substances

  • Cholic Acids
  • Detergents
  • Receptors, Neuropeptide
  • neuropeptide FF receptor
  • 3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate