FEBS Letters | |
Molecular cloning of a novel gene involved in serotonin receptor‐mediated signal transduction in rat stomach | |
Takii, Takemasa1  Ohya, Susumu2  Matsumori, Miki2  Onozaki, Kikuo1  Yamazaki, Hiroko F.2  Imaizumi, Yuji2  Watanabe, Minoru2  | |
[1] Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabedori, Mizuhoku, Nagoya 467, Japan;Department of Chemical Pharmacology, Faculty of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabedori, Mizuhoku, Nagoya 467, Japan | |
关键词: 5-HT receptor; cDNA cloning; Xenopus oocyte; In vitro translation; Northern blotting; [3H]Spiperone binding assay; | |
DOI : 10.1016/S0014-5793(96)01485-8 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
In Xenopus oocytes injected with small size mRNAs (500–700 b), obtained from rat stomach by fractionation, application of 10 μM 5-HT induced a substantial Ca2+-activated Cl− current (ICl-Ca). ICl-Ca was not elicited by 5-HT in native oocytes. Consistent results from this assay in the oocyte expression system motivated cDNA cloning experiments. A novel cDNA (namedattomacherotonin receptor-related cDNA: RSS cDNA) which encodes a small protein involved in specific 5-HT receptor-mediated ICl-Ca activation was identified. The molecular weight of RSS protein in the reticulocyte lysate translation system (∼10 kDa) is identical to that calculated from the amino acid sequence. Computer-aided analysis of the predicted protein does not show any obvious sequence homologies (<18%) to any other proteins including G protein-coupled receptors. Northern analysis revealed that RSS mRNA is ubiquitously expressed at varying levels in a number of different tissues. Furthermore, the binding of [3H]spiperone, a 5-HT2 receptor antagonist, was examined in CHO cells, which highly expressed RSS transcripts (named CHO-RSS). Specific binding of [3H]spiperone was not clearly observed in native CHO but was detected in CHO-RSS. The dissociation constant was 10.3 nM in CHO-RSS. These results suggest that RSS protein may be a factor which facilitates 5-HT receptor expression or, alternatively, an enhancer of the affinity of native 5-HT receptor to 5-HT.
【 授权许可】
Unknown
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