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A generic method for the production of cell lines expressing high levels of 7-transmembrane receptors.

Authors
  • Koller, K J
  • Whitehorn, E A
  • Tate, E
  • Ries, T
  • Aguilar, B
  • Chernov-Rogan, T
  • Davis, A M
  • Dobbs, A
  • Yen, M
  • Barrett, R W
Type
Published Article
Journal
Analytical biochemistry
Publication Date
Jul 15, 1997
Volume
250
Issue
1
Pages
51–60
Identifiers
PMID: 9234898
Source
Medline
License
Unknown

Abstract

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.

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