Renin synthesis by canine aortic smooth muscle cells in culture.
Renin synthesis by canine aortic smooth muscle cells in culture.
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培养物中犬主动脉平滑肌细胞合成肾素。
DOI：
10.1016/0024-3205(82)90641-5
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发表时间：
1982
期刊：
影响因子：
6.1
通讯作者：
Haber,E
中科院分区：
文献类型：
--
作者：
Re,R;Fallon,JT;Dzau,V;Quay,SC;Haber,E
在动脉组织的匀浆中检测到血管紧张素-I生成活性，但尚不清楚这种酶活性是由于肾素本身的存在还是由于其他蛋白酶如组织蛋白酶D的作用。在采用无肾犬血浆作为底物并缓冲至pH 7.4的测定系统中，我们检测到犬主动脉平滑肌细胞匀浆中的血管紧张素-I生成活性。这种酶的活性在很大程度上是可重复的肾素特异性抗血清提出了纯犬肾肾素。培养的动脉平滑肌细胞的免疫荧光研究也进行了使用肾素特异性抗血清。颗粒细胞质免疫荧光检测时，使用特异性抗肾素血清，但不采用免疫前血清。在染色过程中，向肾素抗血清中加入纯犬肾素抑制了颗粒免疫荧光，证实了染色的特异性。最后，进行了生物合成放射性标记研究。免疫沉淀新合成的蛋白质与抗肾素血清和葡萄球菌蛋白A，然后通过凝胶电泳和放射自显影证明合成的免疫反应性蛋白质的分子量的肾素。用纯的犬肾素预处理抗肾素血清，导致该免疫反应蛋白带消失。因此，这些研究提供了多条线索的证据表明，在原位合成的血管平滑肌细胞的肾素。
Angiotensin-I generating activity has been detected in homogenates of arterial tissue but it remains unclear whether this enzymatic activity results from the presence of renin itself or from the action of other proteases such as cathepsin D. In an assay system employing anephric dog plasma as substrate and buffered to pH 7.4, we detected angiotensin-I generating activity in homogenates of canine aortic smooth muscle cells. This enzymatic activity was in large part inhibitable by renin-specific antisera raised to pure canine renal renin. Immunofluorescent study of cultured arterial smooth muscle cells was also performed using renin specific antiserum. Granular cytoplasmic immunofluorescence was detected when specific antirenin serum was used but not when preimmune serum was employed. The addition of pure canine renin to the renin antiserum during staining suppressed the granular immunofluorescence confirming the specificity of staining. Finally, biosynthetic radiolabelling studies were performed. Immunoprecipitation of newly synthesized proteins with antirenin serum and staphylococcal protein A followed by gel electrophoresis and autoradiography demonstrated the synthesis of an immunoreactive protein with the molecular weight of renin. Pretreatment of the antirenin serum with pure canine renin resulted in the disappearance of this immunoreactive protein band. Thus these studies provide multiple lines of evidence to indicate the in situ synthesis of renin by vascular smooth muscle cells.