Hirasawa A, Shibata K, Kotosai K, Tsujimoto G
Department of Molecular Cell Pharmacology, National Children's Medical Research Center, Tokyo, Japan.
Biochem Biophys Res Commun. 1994 Aug 30;203(1):72-9. doi: 10.1006/bbrc.1994.2150.
We have cloned a human vasopressin receptor from human mesenteric artery using RACE (Rapid Amplification of cDNA Ends) methods. The deduced amino acid sequence of the clone (HV-RACE) encodes a protein of 418 amino acids that showed a strong sequence homology to the previously cloned rat V1A vasopressin receptor. The [3H] arginine vasopressin (AVP) binding to HV-RACE expressed in COS-7 cells was potently inhibited by AVP (Ki = 2.9 nM). Interestingly, a new non-peptide "V1-selective" antagonist OPC-21268 exhibited markedly higher affinity for rat V1A receptor (Ki = 57 nM) rather than for HV-RACE (Ki = 56 microM). With the reverse-transcription polymerase chain reaction assay, we observed a large amount of HV-RACE transcripts in the mesenteric artery, while a small amount in a variety of other tissues. The data show that the clone HV-RACE encodes a human vascular-type vasopressin receptor cDNA.
我们使用cDNA末端快速扩增（RACE）方法从人肠系膜动脉中克隆了一种人血管加压素受体。该克隆（HV-RACE）推导的氨基酸序列编码一个由418个氨基酸组成的蛋白质，它与先前克隆的大鼠V1A血管加压素受体显示出很强的序列同源性。在COS-7细胞中表达的HV-RACE与[3H]精氨酸血管加压素（AVP）的结合被AVP强烈抑制（Ki = 2.9 nM）。有趣的是，一种新的非肽类“V1选择性”拮抗剂OPC-21268对大鼠V1A受体（Ki = 57 nM）的亲和力明显高于对HV-RACE（Ki = 56 μM）的亲和力。通过逆转录聚合酶链反应分析，我们在肠系膜动脉中观察到大量的HV-RACE转录本，而在其他多种组织中含量较少。数据表明，克隆HV-RACE编码一种人血管型血管加压素受体cDNA。
