Biochemical similarity of expressed human prorenin and native inactive renin.
Biochemical similarity of expressed human prorenin and native inactive renin.
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表达的人肾素原和天然无活性肾素的生化相似性。
DOI：
10.1161/01.hyp.8.6_pt_2.ii78
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Fritz,LC
中科院分区：
文献类型：
--
作者：
Hsueh,WA;Do,YS;Shinagawa,T;Tam,H;Ponte,PA;Baxter,JD;Shine,J;Fritz,LC
前肾素由用人前肾素表达构建体转染的哺乳动物细胞分泌。本研究的目的是比较在培养基中表达的原肾素的理化性质与已知的人非活性肾素的特征，其占血浆和肾脏中近一半的肾素。我们发现，表达的人前肾素非常类似于人肾和血浆中的非活性肾素。表达的前肾素是无活性的，可以同样激活酸（透析至pH 3.3）或胰蛋白酶。酸激活是完全可逆的，再次暴露于酸可以重新激活表达的失活的肾素。暴露于寒冷（-5 ℃持续3天）也可以激活表达的肾素。以绵羊为底物的酸活化表达的肾素的米氏常数为0.29 μ M，最适pH为7.8。表达的失活的肾素结合到cibacron蓝亲和柱上，并且可以用0.5M NaCl洗脱。所有上述特征类似于人肾和血浆中的无活性肾素。此外，表达的原肾素和人绒毛膜肾素的分子量为47，000，如通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定的，和46，000，如通过高效液相色谱测定的。这些数据，结合已发表的观察结果，即天然人非活性肾素与针对肾素前片段中氨基酸序列产生的抗体交叉反应，为人类非活性肾素是前肾素的假设提供了强有力的支持。
Prorenin is secreted by mammalian cells transfected with a human preprorenin expression construct. The purpose of this investigation was to compare the physicochemical properties of expressed prorenin in culture medium with the known characteristics of human inactive renin, which accounts for nearly half the renin in plasma and kidney. We found that expressed human prorenin strongly resembles human renal and plasma inactive renin. The expressed prorenin was inactive and could be equally activated by acid (dialysis to pH 3.3) or trypsin. Acid activation was completely reversible; reexposure to acid could reactivate the expressed inactive renin. Exposure to cold (-5 degrees C for 3 days) could also activate expressed renin. The Michaelis-Menten constant of acid-activated expressed renin with sheep substrate was 0.29 microM, and the pH optimum was 7.8. Expressed inactive renin bound to a cibacron-blue affinity column and could be eluted with 0.5M NaCl. All the above characteristics resemble those of human renal and plasma inactive renin. In addition, the molecular weight of expressed prorenin and human chorionic renin was 47,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 46,000, as measured by high-performance liquid chromatography. These data, taken together with the published observation that native human inactive renin cross-reacts with antibodies generated against amino acid sequences in the prosegment of renin, provide strong support for the hypothesis that human inactive renin is prorenin.