A Generic Method for the Production of Cell Lines Expressing High Levels of 7-Transmembrane Receptors

✍ Scribed by Kerry J. Koller; Erik A. Whitehorn; Emily Tate; Tracy Ries; Barbara Aguilar; Tania Chernov-Rogan; Ann M. Davis; Andrea Dobbs; Mary Yen; Ronald W. Barrett

• Book ID: 102566462
• Publisher: Elsevier Science
• Year: 1997
• Tongue: English
• Weight: 215 KB
• Volume: 250
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1997.2190

✦ Synopsis

G-protein-coupled or 7-transmembrane receptors (7TMRs) are often studied after heterologous expression in mammalian cells such as COS-7, CHO-K1, or HEK-293s. In this paper, we describe the development of a rapid and generic method for producing stable Chinese hamster ovary cell lines expressing high levels of recombinant 7TMRs by N-terminal tagging these proteins with the hemagglutinin (HA) sequence. To illustrate the broad applicability of this technique, we have presented data from cell lines expressing a glycoprotein hormone receptor for follicle-stimulating hormone (FSHR), CXC- (CXCR-2), and CC-chemokine (CCR-1) receptors and peptide receptors from the somatostatin (SSTR1, 2, 5) and neuropeptide Y (NPY-Y2, -Y4 Rs) families. Typically, cell lines with a receptor 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.