Dubin D, Pratt R E, Hui K Y, Dzau V J
Falk Cardiovascular Research Center, Stanford, CA 94305-5246.
J Hypertens. 1991 Jun;9(6):483-6. doi: 10.1097/00004872-199106000-00001.
Human renin is synthesized as an inactive zymogen (prorenin) which is processed to the active form. We synthesized an 11-amino acid peptide which spans the human prorenin processing site in order to develop a simple assay to study human prorenin activation. Six enzymes which are capable of activating recombinant prorenin in vitro were studied. Four of these enzymes digested the synthetic peptide in a specific fashion, as analyzed by reverse-phase high-performance liquid chromatography. Amino acid analysis of the purified digestion products revealed that trypsin cleaves between Arg-Leu, the authentic processing site, while kallikrein, plasmin and elastase all cleaved at alternate sites. On the other hand, pepsin and cathepsin D did not cleave this substrate, suggesting that the activation of prorenin by these proteases might occur at a site distinct from the authentic processing site. Our data suggest that this synthetic peptide may be used as a simple and specific assay for prorenin activation.
人肾素以无活性的酶原（前肾素）形式合成，该酶原会被加工成活性形式。我们合成了一种跨越人前肾素加工位点的11个氨基酸的肽，以便开发一种简单的检测方法来研究人前肾素的激活。研究了六种能够在体外激活重组前肾素的酶。通过反相高效液相色谱分析，其中四种酶以特定方式消化合成肽。对纯化的消化产物进行氨基酸分析表明，胰蛋白酶在精氨酸-亮氨酸之间切割，这是真正的加工位点，而激肽释放酶、纤溶酶和弹性蛋白酶均在其他位点切割。另一方面，胃蛋白酶和组织蛋白酶D不会切割该底物，这表明这些蛋白酶对前肾素的激活可能发生在与真正加工位点不同的位点。我们的数据表明，这种合成肽可作为一种简单且特异的前肾素激活检测方法。
