THE HUMAN ENDOTHELIN FAMILY - 3 STRUCTURALLY AND PHARMACOLOGICALLY DISTINCT ISOPEPTIDES PREDICTED BY 3 SEPARATE GENES
THE HUMAN ENDOTHELIN FAMILY - 3 STRUCTURALLY AND PHARMACOLOGICALLY DISTINCT ISOPEPTIDES PREDICTED BY 3 SEPARATE GENES
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DOI：
10.1073/pnas.86.8.2863
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发表时间：
1989-04-01
影响因子：
11.1
通讯作者：
MASAKI, T
中科院分区：
文献类型：
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作者：
INOUE, A;YANAGISAWA, M;MASAKI, T
通过用编码内皮素序列的一部分的合成寡核苷酸探针在低杂交严格性下筛选基因组DNA文库，克隆了三个不同的人内皮素相关基因。用相同的寡核苷酸探针进行基因组Southern印迹分析，不仅在人类基因组中，而且在猪和大鼠基因组中显示了三个相应的染色体位点。这三个人基因的核苷酸序列在编码21个残基（成熟）内皮素的区域内高度保守，尽管编码前肽的一部分的紧邻上游外显子序列几乎没有相似性。此外，每个人类基因预测了一个假定的21个残基的肽，类似但彼此不同：（i）“经典”内皮素（ET-1），（ii）[Trp 6，Leu 7]内皮素（ET-2），和（iii）[Thr 2，Phe 4，Thr 5，Tyr 6，Lys 7 Tyr 14]内皮素（ET-3）。根据推导的氨基酸序列合成了ET-1、ET-2和ET-3，通过收缩离体猪冠状动脉条和静脉注射麻醉大鼠来测定其生物活性。这些合成肽均产生强的缩血管和升压反应。然而，药理活性的定量分布有很大的不同，这三个异肽，提示可能存在内皮素受体亚型。
Three distinct human endothelin-related genes were cloned by screening a genomic DNA library under a low hybridization stringency with a synthetic oligonucleotide probe encoding a portion of the endothelin sequence. Genomic Southern blot analysis with the same oligonucleotide probe showed three corresponding chromosomal loci not only in the human genome but also in porcine and rat genomes. The nucleotide sequences of the three human genes were highly conserved within the regions encoding the 21-residue (mature) endothelins, in spite of the fact that the immediately upstream exon sequences, which encode a part of the propeptides, retained little similarity. Moreover, each of the human genes predicted a putative 21-residue peptide, similar to but distinct from each other: (i) the "classical" endothelin (ET-1), (ii) [Trp6,Leu7]endothelin (ET-2), and (iii) [Thr2,Phe4,Thr5,Tyr6,Lys7Tyr14]endothelin (ET-3). Synthetic ET-1, ET-2, and ET-3 were prepared according to the deduced amino acid sequences, and the biological activities were assayed by contraction of isolated porcine coronary artery strips and by intravenous injection to anesthetized rats. All these synthetic peptides produced strong vasoconstrictor and pressor responses. However, the quantitative profiles of the pharmacological activities were considerably different among the three isopeptides, suggesting the possible existence of endothelin receptor subtypes.