Retrovirus-Mediated Expression of the Galr1 Galanin Receptor: Implication for Efficient Stable Expression of Functional G Protein-Coupled Receptors

Abstract

The rat GalR1 galanin receptor was used as a prototypic G protein-coupled receptor to test the feasibility of heterologous expression in a retrovirus-based system. The system utilizes an independent retroviral vector pMX, a virus-packaging cell line BOSC23 and a pre-B cell line BA/F3 as the host for expression. A polyclonal cell population that expresses high ligand affinity (KD = 0.18 nM) and high level (7 pmol/mg) of GalR1 was generated within days with no drug sensitivity-based selection. The expression represented a 20-fold increase over the expression level of GalR1 achieved in CHO cells. The affinity of galanin for the expressed receptor was decreased by 19-fold in the presence of GTP-gamma-S, suggesting that the expression system can produce active galanin receptor functionally coupled to G proteins. The fast and efficient method to generate stable cell lines and to prepared large quantities of receptors may provide a general application for expression of other G protein-coupled receptors.

Galanin and galanin‐like peptide modulate neurite outgrowth via protein kinase C‐mediated activation of extracellular signal‐related kinase

“…The similarity between the ability of galanin and GALP to activate ERK suggests that GalR2 may play a predominant role in ERK activation in PC12 cells. This is consistent with previous reports that GalR2 mediates ERK activation in non‐neuronal cells (Wang et al. , 1998).…”

“…, 2000). Overexpression of GalR1 and GalR2 in Chinese hamster ovary cells leads to galanin‐induced ERK phosphorylation (Wang et al. , 1998).…”