| RRC ID |
42295
|
| 著者 |
Yin H, Ukena K, Ubuka T, Tsutsui K.
|
| タイトル |
A novel G protein-coupled receptor for gonadotropin-inhibitory hormone in the Japanese quail (Coturnix japonica): identification, expression and binding activity.
|
| ジャーナル |
J Endocrinol
|
| Abstract |
We recently identified a novel hypothalamic dodecapeptide inhibiting gonadotropin release in the Japanese quail (Coturnix japonica). This novel peptide was therefore named gonadotropin-inhibitory hormone (GnIH). The GnIH precursor encoded one GnIH and two GnIH-related peptides (GnIH-RP-1 and GnIH-RP-2) that shared the same C-terminal motif, Leu-Pro-Xaa-Arg-Phe-NH(2) (Xaa=Leu or Gln; LPXRF-amide peptides). Identification of the receptor for GnIH is crucial to elucidate the mode of action of GnIH. We therefore identified the receptor for GnIH in the quail diencephalon and characterized its expression and binding activity. We first cloned a cDNA encoding a putative GnIH receptor by a combination of 3' and 5' rapid amplification of cDNA ends (RACE) using PCR primers designed from the sequence for the receptor for rat RF-amide-related peptide (RFRP), an orthologous peptide of GnIH. Hydrophobic analysis revealed that the putative GnIH receptor possessed seven transmembrane domains, indicating a new member of the G protein-coupled receptor superfamily. The crude membrane fraction of COS-7 cells transfected with the putative GnIH receptor cDNA specifically bound to GnIH and GnIH-RPs in a concentration-dependent manner. Scatchard plot analysis of the binding showed that the identified GnIH receptor possessed a single class of high-affinity binding sites (K(d)=0.752 nM, B(max)=24.8 fmol/mg protein). Southern blotting analysis of reverse transcriptase-mediated PCR products revealed the expression of GnIH receptor mRNA in the pituitary gland and several brain regions including diencephalon in the quail. These results suggest that GnIH acts directly on the pituitary via GnIH receptor to inhibit gonadotropin release. GnIH may also act on the hypothalamus to inhibit gonadotropin-releasing hormone release.
|
| 巻・号 |
184(1)
|
| ページ |
257-66
|
| 公開日 |
2005-1-1
|
| DOI |
10.1677/joe.1.05926
|
| PII |
184/1/257
|
| PMID |
15642802
|
| MeSH |
Amino Acid Sequence
Animals
Avian Proteins / metabolism
Base Sequence
Binding Sites
Blotting, Southern
COS Cells
Coturnix
Humans
Hypothalamic Hormones / metabolism
Male
Molecular Sequence Data
Protein Binding
RNA, Messenger / analysis*
Rats
Receptors, G-Protein-Coupled / genetics*
Receptors, G-Protein-Coupled / metabolism
Sequence Homology
Transfection / methods
|
| IF |
4.041
|
| 引用数 |
156
|
|
WOS 分野
|
ENDOCRINOLOGY & METABOLISM
|
| リソース情報 |
| ヒト・動物細胞 |
COS-7(RCB0539) |
