Kohzuki M, Abe K, Yasujima M, Yoshinaga K
Second Department of Internal Medicine, Tohoku University School of Medicine, Sendai, Japan.
Nihon Naibunpi Gakkai Zasshi. 1991 Aug 20;67(8):849-60. doi: 10.1507/endocrine1927.67.8_849.
Sarafotoxin S6B(SRT), a peptide in snake venom, has a high degree of sequence homology with endothelin (ET) and both are potent vasoconstrictors. In order to determine whether SRT acts via the ET receptor, we performed autoradiographic binding studies on rat tissues using the radioligands, 125I-ET-1 and 125I-SRT and computerized in vitro autoradiography. In the heart, a high density of ET binding was found in the atria, and moderate density was found in the ventricles. A high density of ET-1 was found in the heart. In the kidney, ET-1 binding occurred in association with glomeruli, outer cortex, and inner stripe and inner medulla. In the adrenal, a high density of ET-1 binding occurred in the medulla as well as the zona glomerulosa. The binding affinity constant (KA) for ET-1 binding in these sites ranged from 1-10 x 10(9) M-1. Although SRT was 5-100 fold weaker than ET-1 in displacing 125I-ET-1 from these sites, 1 microM unlabelled SRT completely abolished 125I-ET-1 binding in all sites. Other venom peptides or unrelated peptides did not affect 125I-ET-1 binding. Moreover, the pattern of 125I-SRT binding in rat tissues by in vitro autoradiography was identical with that of 125I-ET-1 binding, and both unlabelled SRT and unlabelled ET-1 fully competed with 125I-SRT for binding. These results provide evidence that SRT binds to the ET binding sites in a range of rat tissues.
萨拉弗毒素S6B(SRT)是一种蛇毒中的肽,与内皮素(ET)具有高度的序列同源性,两者都是强效血管收缩剂。为了确定SRT是否通过ET受体发挥作用,我们使用放射性配体125I-ET-1和125I-SRT以及计算机体外放射自显影技术对大鼠组织进行了放射自显影结合研究。在心脏中,心房发现高密度的ET结合,心室发现中等密度的ET结合。在心脏中发现高密度的ET-1。在肾脏中,ET-1结合发生在肾小球、外皮质、内带和内髓质。在肾上腺中,髓质以及球状带发现高密度的ET-1结合。这些位点中ET-1结合的结合亲和力常数(KA)范围为1-10×10(9)M-1。尽管SRT在从这些位点置换125I-ET-1方面比ET-1弱5-100倍,但1μM未标记的SRT完全消除了所有位点的125I-ET-1结合。其他蛇毒肽或无关肽不影响125I-ET-1结合。此外,通过体外放射自显影在大鼠组织中125I-SRT结合的模式与125I-ET-1结合的模式相同,未标记的SRT和未标记的ET-1都与125I-SRT完全竞争结合。这些结果提供了证据表明SRT在一系列大鼠组织中与ET结合位点结合。