Isolation of an angiotensin II-binding protein from liver.
Isolation of an angiotensin II-binding protein from liver.
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从肝脏中分离血管紧张素 II 结合蛋白。
DOI：
10.1073/pnas.81.6.1679
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发表时间：
1984
影响因子：
11.1
通讯作者：
Soffer,RL
中科院分区：
文献类型：
--
作者：
Sen,I;Bull,HG;Soffer,RL
一种特异性结合血管紧张素II的蛋白质，在用毛地黄皂苷处理从兔肝颗粒中溶解后，通过两种独立的方法以几乎均质的形式分离出来。通过这些方法中的任一种纯化的蛋白质在大小上类似于通过使用辛二酸二琥珀酰亚胺酯使放射性碘化的血管紧张素II与其在溶解的提取物中的受体交联而制备的单个放射性大分子组分。在第一种技术中，血管紧张素II作为亲和配体，从已去除血管紧张素降解活性的制剂中特异性提取蛋白质。在第二种方法中，血管紧张素II-蛋白复合物被抗血管紧张素II抗体和葡萄球菌蛋白A-Sepharose特异性沉淀。可以用血管紧张素II或4 M MgCl 2从亲和柱上洗脱蛋白质。血管紧张素II-蛋白质复合物在含巯基试剂存在下解离，因此这些试剂可用于将其从化学或免疫亲和基质中分离出来。含巯基试剂的这种效应以及变性后分离的蛋白质在其还原形式下表现出比在其未还原形式下更慢的电泳迁移率的矛盾观察结果表明，该蛋白质的结合构型可能对还原敏感。
A protein that specifically binds angiotensin II has been isolated in nearly homogeneous form by two independent approaches after solubilization from rabbit liver particles by treatment with digitonin. The protein purified by either of these methods resembles in size the single radioactive macromolecular component made by using disuccinimidyl suberate to crosslink radioiodinated angiotensin II with its receptor in the solubilized extract. In the first technique, angiotensin II as an affinity ligand specifically extracted the protein from a preparation that had been freed of angiotensin-degrading activity. In the second approach, the angiotensin II-protein complex was specifically precipitated by anti-angiotensin II antibodies and staphylococcal protein A-Sepharose. The protein could be eluted from the affinity column with angiotensin II or 4 M MgCl2. The angiotensin II-protein complex dissociated in the presence of sulfhydryl-containing reagents, and these could therefore be used to elute it from either the chemical or the immunoaffinity-based matrix. This effect of sulfhydryl-containing reagents and the paradoxical observation that the isolated protein after denaturation exhibited a slower electrophoretic mobility in its reduced form that in its unreduced form suggest that the binding configuration of this protein may be sensitive to reduction.