The Mouse GalR2 Galanin Receptor: Genomic Organization, cDNA Cloning, and Functional Characterization |
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Authors: | Ling Pang Tanaz Hashemi Hu-Jung J. Lee Maureen Maguire Michael P. Graziano Marvin Bayne Brian Hawes Gwendolyn Wong Suke Wang |
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Affiliation: | Department of CNS/CV Biological Research, Schering-Plough Research Institute, Kenilworth, New Jersey, U.S.A. |
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Abstract: | Abstract: The diverse physiological actions of galanin are thought to be mediated through activation of galanin receptors (GalRs). We report the genomic and cDNA cloning of a mouse GalR that possesses a genomic structure distinct from that of GalR1 and encodes a functional galanin receptor. The mouse GalR gene consists of two exons separated by a single intron within the protein-coding region. The splicing site for the intron is located at the junction between the third transmembrane domain and the second intracellular loop. The cDNA encodes a 370-amino acid putative G protein-coupled receptor that is markedly different from human GalR1 and rat GalR3 (38 and 57%) but shares high homology with rat GalR2 (94%). In binding studies utilizing membranes from COS-7 cells transfected with mouse GalR2 cDNA, the receptor displayed high affinity ( K D = 0.47 n M ) and saturable binding with 125I-galanin ( B max = 670 fmol/mg). The radioligand binding can be displaced by galanin and its analogues in a rank order: galanin ⋍ M40 ⋍ M15 ⋍ M35 ⋍ C7 ⋍ galanin (2–29) ⋍ galanin (1–16) ≫ galanin (10–29) ⋍ galanin (3–29), which resembles the pharmacological profile of the rat GalR2. Receptor activation by galanin in COS-7 cells stimulated phosphoinositide metabolism, which was not reversed by pertussis toxin. Thus, the galanin receptor encoded in the cloned mouse GalR gene is the type 2 galanin receptor and is active in both ligand binding and signaling assays. |
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Keywords: | Galanin Galanin receptors Ligand binding Gene organization Pharmacological profile Intracellular signaling |
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