Cloning and characterization of a human orphan family C G-protein coupled receptor GPRC5D
Research output: Contribution to journal › Journal article › Research › peer-review
Recently three orphan G-protein coupled receptors, RAIG1, GPRC5B and GPRC5C, with homology to members of family C (metabotropic glutamate receptor-like) have been identified. Using the protein sequences of these receptors as queries we identified overlapping expressed sequence tags which were predicted to encode an additional subtype. The full length coding regions of mouse mGprc5d and human GPRC5D were cloned and shown to contain predicted open reading frames of 300 and 345 amino acids, respectively. GPRC5D has seven putative transmembrane segments and is expressed in the cell membrane. The four human receptor subtypes, which we assign to group 5 of family C GPCRs, show 31-42% amino acid sequence identity to each other and 20-25% sequence identity to the transmembrane domains of metabotropic glutamate receptor subtypes 2 and 3 and other family C members. In contrast to the remaining family C members, the group 5 receptors have short amino terminal domains of some 30-50 amino acids. GPRC5D was shown to be clustered with RAIG1 on chromosome 12p13.3 and like RAIG1 and GPRC5B to consist of three exons, the first exon being the largest containing all seven transmembrane segments. GPRC5D mRNA is widely expressed in the peripheral system but all four receptors show distinct expression patterns. Interestingly, mRNA levels of all four group 5 receptors were found in medium to high levels in the kidney, pancreas and prostate and in low to medium levels in the colon and the small intestine, whereas other organs only express a subset of the genes. In an attempt to delineate the signal transduction pathway(s) of the orphan receptors, a series of chimeric receptors containing the amino terminal domain of the calcium sensing receptor or metabotropic glutamate receptor subtype 1, and the seven transmembrane domain of the orphan receptors were constructed and tested in binding and functional assays.
|Journal||BBA General Subjects|
|Number of pages||12|
|Publication status||Published - 2001|
- Amino Acid Sequence, Animals, Base Sequence, Cell Membrane, Cloning, Molecular, GTP-Binding Proteins, Gene Library, Green Fluorescent Proteins, Humans, Kidney, Luminescent Proteins, Male, Mice, Molecular Sequence Data, Pancreas, Prostate, Quisqualic Acid, Receptors, Cell Surface, Receptors, G-Protein-Coupled, Receptors, Metabotropic Glutamate, Recombinant Fusion Proteins, Sequence Alignment, Sequence Homology, Signal Transduction, Transfection, Tretinoin, Tumor Cells, Cultured