Molecular cloning and characterisation of GPR74 a novel G-protein coupled receptor closest related to the Y-receptor family
| Autor: | Debra J. Gilbert, Joanna Crawford, Marjorie Liu, Grant R. Sutherland, Michelle Couzens, Rachel M. C. Parker, Neal G. Copeland, Nancy A. Jenkins, Helen J. Eyre, Herbert Herzog |
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| Rok vydání: | 2000 |
| Předmět: |
Male
Receptors Neuropeptide Receptors Cell Surface Molecular cloning Biology Ligands Homology (biology) Substrate Specificity Cellular and Molecular Neuroscience Exon Mice Complementary DNA Animals Humans Amino Acid Sequence RNA Messenger Cloning Molecular Molecular Biology Gene G protein-coupled receptor Expressed Sequence Tags Expressed sequence tag Base Sequence cDNA library Myocardium Brain Chromosome Mapping Exons Molecular biology Heterotrimeric GTP-Binding Proteins Introns Receptors Neuropeptide Y Organ Specificity Female Chromosomes Human Pair 4 Sequence Alignment |
| Zdroj: | Brain research. Molecular brain research. 77(2) |
| ISSN: | 0169-328X |
| Popis: | A novel gene product, GPR74, with homology to the seven transmembrane-domain receptor superfamily, has been cloned. GPR74 has been identified from the expressed sequence tags (EST) database. Subsequent PCR amplification of that sequence and screening of a human heart cDNA library led to the isolation of a 1.7-kb cDNA clone encoding a protein of 408 amino acids. GPR74 shows highest amino acid identity (33%) to the human neuropeptide Y-receptor subtype Y2. The human and mouse genes for GPR74 have been isolated and their exon–intron structures determined. In both species the gene consists of four exons spanning around 20 kb with the exon–intron borders being 100% conserved. Northern analysis of various human tissues reveals highest levels of mRNA expression in brain and heart. In situ hybridisation analysis of rat brain tissue confirms this result and identifies the hippocampus and amygdala nuclei as the brain areas with particular high expression of GPR74 mRNA. Fluorescence in situ hybridisation, PCR analysis on a radiation hybrid panel and interspecific mouse backcross mapping have localised the genes to human chromosome 4q21 and mouse chromosome 5. Expression of the human GPR74 cDNA as a GFP-fusion protein in various cell lines reveals the inability of the recombinant receptor protein to reach the cell surface. This is consistent with the lack of NPY specific binding in these cells and suggests that unknown factors are required for a full functional receptor complex. |
| Databáze: | OpenAIRE |
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