Cloning and characterization of a human orphan family C G-protein coupled receptor GPRC5D
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Cloning and characterization of a human orphan family C G-protein coupled receptor GPRC5D. / Bräuner-Osborne, H; Jensen, A A; Sheppard, P O; Brodin, Birger; Krogsgaard-Larsen, P; O'Hara, P.
In: BBA General Subjects, Vol. 1518, No. 3, 2001, p. 237-48.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Cloning and characterization of a human orphan family C G-protein coupled receptor GPRC5D
AU - Bräuner-Osborne, H
AU - Jensen, A A
AU - Sheppard, P O
AU - Brodin, Birger
AU - Krogsgaard-Larsen, P
AU - O'Hara, P
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
KW - Amino Acid Sequence
KW - Animals
KW - Base Sequence
KW - Cell Membrane
KW - Cloning, Molecular
KW - GTP-Binding Proteins
KW - Gene Library
KW - Green Fluorescent Proteins
KW - Humans
KW - Kidney
KW - Luminescent Proteins
KW - Male
KW - Mice
KW - Molecular Sequence Data
KW - Pancreas
KW - Prostate
KW - Quisqualic Acid
KW - Receptors, Cell Surface
KW - Receptors, G-Protein-Coupled
KW - Receptors, Metabotropic Glutamate
KW - Recombinant Fusion Proteins
KW - Sequence Alignment
KW - Sequence Homology
KW - Signal Transduction
KW - Transfection
KW - Tretinoin
KW - Tumor Cells, Cultured
M3 - Journal article
C2 - 11311935
VL - 1518
SP - 237
EP - 248
JO - B B A - General Subjects
JF - B B A - General Subjects
SN - 0304-4165
IS - 3
ER -
ID: 37899786