Solubilization and functional reconstitution of human neuropeptide FF₂ receptors

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• 作者：Frank Talmont ; Catherine Mollereau ; Isabelle Muller ; Jean-Marie Zajac
• 关键词：Neuropeptide FF ; Receptor ; Solubilization ; Reconstitution ; Binding sites
• 刊名：Analytical Biochemistry
• 出版年：2010
• 出版时间：15 March 2010
• 年：2010
• 卷：398
• 期：2
• 页码：225-229
• 全文大小：185 K

文摘

Neuropeptide FF (NPFF, FLFQPQRFamide) receptors modulate endogenous opioid functions. Here, we report the solubilization of the human NPFF₂ 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.