A GENERIC METHOD FOR THE PRODUCTION OF CELL-LINES EXPRESSING HIGH-LEVELS OF 7-TRANSMEMBRANE RECEPTORS

G-protein-coupled or 7-transmembrane receptors (7TMRs) are often studi ed after heterologous expression in mammalian cells such as COS-7, CHO -K1, or HEK-293s. In this paper, we describe the development of a rapi d and generic method for producing stable Chinese hamster ovary cell l ines expressing high levels of recombinant 7TMRs by N-terminal tagging these proteins with the hemagglutinin (HA) sequence. To illustrate th e broad applicability of this technique, we have presented data from c ell lines expressing a glycoprotein hormone receptor for follicle-stim ulating hormone (FSHR), CXC- (CXCR-2), and CC-chemokine (CCR-1) recept ors and peptide receptors from the somatostatin (SSTR1, 2, 5) and neur opeptide Y (NPY-Y2, -Y4 Rs) families. Typically, cell lines with a rec eptor density of 1 to 15 pmol/mg protein are produced with this method . The presence of the HA tag does not adversely affect the binding or functional activity of the receptors. (C) 1997 Academic Press.