Shinagawa T, Do Y S, Tam H, Hsueh W A
Biochem Biophys Res Commun. 1986 Sep 14;139(2):446-54. doi: 10.1016/s0006-291x(86)80011-0.
Renin was completely purified from human kidney cortex using a rapid 3-step procedure which included homogenization and ammonium sulfate precipitation, aminohexyl pepstatin affinity chromatography, and affinity chromatography using a synthetic octapeptide renin inhibitor (H-77) with a reduced peptide bond between Leu5 - Leu6. Three kg of cortex dissected from 10 kg of human cadaver kidney yielded 0.7 mg protein with a specific activity of 1123 GU/mg protein and an overall recovery of 52%. Both gel filtration high pressure liquid chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) revealed a molecular weight of 44,000, although 22,000 and 18,000 molecular weight bands were also identified by SDS PAGE. Amino terminal sequencing demonstrated a leucine residue at the 1 position indicating that prorenin is converted to renin following cleavage at the carboxyl end of two dibasic residues, Lys-2-Arg-1. Sequencing of the first 19 amino acids was in agreement with the sequence deduced from human renin cDNA sequence.
采用一种快速的三步法从人肾皮质中完全纯化肾素，该方法包括匀浆和硫酸铵沉淀、氨基己基胃蛋白酶抑制剂亲和色谱以及使用在Leu5 - Leu6之间肽键减少的合成八肽肾素抑制剂（H - 77）的亲和色谱。从10千克人尸体肾脏中切取的3千克皮质得到0.7毫克蛋白质，比活性为1123酶活力单位/毫克蛋白质，总回收率为52%。凝胶过滤高压液相色谱和十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS - PAGE）均显示分子量为44,000，不过SDS - PAGE也鉴定出了分子量为22,000和18,000的条带。氨基末端测序表明第1位是亮氨酸残基，这表明在两个碱性二肽残基Lys - 2 - Arg - 1的羧基末端裂解后，肾素原转化为肾素。前19个氨基酸的测序结果与从人肾素cDNA序列推导的序列一致。
