Molecular characterization, pharmacological properties and chromosomal localization of the human GALR2 galanin receptor.
The neuropeptide galanin mediates a diverse spectrum of biological activities by interacting with specific G protein-coupled receptors. We have used homology genomic library screening and polymerase chain reaction (PCR) techniques to isolate both genomic and cDNA clones encoding the human homolog of the recently cloned rat GALR2 galanin receptor. By fluorescence in situ hybridization, the gene encoding human GALR2 (GALNR2) has been localized to chromosome 17q25.3. The two coding exons of the human GALNR2 gene, interrupted by an intron positioned at the end of transmembrane domain III, encode a 387 amino acid G protein-coupled receptor with 87% overall amino acid identity with rat GALR2. In HEK-293 cells stably expressing human GALR2, binding of [125I]porcine galanin is saturable and can be displaced by galanin, amino-terminal galanin fragments and chimeric galanin peptides but not by carboxy-terminal galanin fragments. In HEK-293 cells, human GALR2 couples both to Galphaq/11 to stimulate phospholipase C and increase intracellular calcium levels and to Galphai/o to inhibit forskolin-stimulated intracellular cAMP accumulation. A wide tissue distribution is observed by reverse transcriptase (RT)-PCR analysis, with human GALR2 mRNA being detected in many areas of the human central nervous system as well as in peripheral tissues.
Neuroscience Drug Discovery, Bristol-Myers Squibb Pharmaceutical Research Institute, 5 Research Parkway, Wallingford, CT 06492, USA. fathi@bms.com
Z
Z
Fathi
P M
PM
Battaglino
L G
LG
Iben
H
H
Li
E
E
Baker
D
D
Zhang
R
R
McGovern
C D
CD
Mahle
G R
GR
Sutherland
T P
TP
Iismaa
K E
KE
Dickinson
I A
IA
Zimanyi
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002874:Chromosome Mapping; D002886:Chromosomes, Human, Pair 17; D019976:Cloning, Organism; D018076:DNA, Complementary; D019204:GTP-Binding Proteins / Q000378:metabolism*; D019004:Galanin / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007700:Kinetics; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000378:metabolism; D051381:Rats; D044088:Receptors, Galanin; D018013:Receptors, Neuropeptide / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013552:Swine; D014162:Transfection
D018076:DNA, Complementary; D010716:Phosphatidylinositols; D044088:Receptors, Galanin; D018013:Receptors, Neuropeptide; D011994:Recombinant Proteins; D019004:Galanin; D019204:GTP-Binding Proteins