Kizuki K, Shimamoto Y, Ikekita M, Moriya H
Adv Exp Med Biol. 1986;198 Pt A:329-37. doi: 10.1007/978-1-4684-5143-6_45.
An inactive form of human urinary kallikrein (inactive HUK) was highly purified from fresh urine collected from healthy men. Inactive HUK was separated from the active kallikrein (HUK) initially presents in the urine by affinity chromatography on a column of aprotinin immobilized on Sepharose 4B and further purified by gel filtration, ion-exchange chromatography and immunoaffinity chromatography on an anti-HUK antibody immobilized Sepharose 4B column. Inactive HUK was rapidly activated by a trace amount of trypsin. While, plasmin, urokinase, thrombin and chymotrypsin caused no activation of inactive HUK. The molecular weights of inactive HUK and HUK were estimated to be 4.8 X 10(4) and 4.5 X 10(4), respectively. The molecular weight of active HUK generated from inactive HUK by the action of trypsin (HUK'') was almost the same as that of HUK. The mobility of inactive HUK was slightly slower than that of HUK on both immunoelectrophoresis and polyacrylamide gel disc electrophoresis. On the other hand, the electrophoretic mobility of HUKK'' was almost the same as that of HUK. These two types of active HUK had no significant difference in the Km values for H-Pro-Phe-Arg-MCA hydrolysis and inhibition profiles by various protease inhibitors and anti-HUK antibody. Inactive HUK was unable to be measured by the direct radioimmunoassay (RIA) but HUK" generated by the action of trypsin could be measured by the RIA.
从健康男性的新鲜尿液中高度纯化出一种人尿激肽释放酶的无活性形式（无活性HUK）。通过固定在琼脂糖4B上的抑肽酶柱进行亲和层析，将无活性HUK与最初存在于尿液中的活性激肽释放酶（HUK）分离，然后通过凝胶过滤、离子交换层析以及固定在琼脂糖4B柱上的抗HUK抗体进行免疫亲和层析进一步纯化。微量胰蛋白酶能迅速激活无活性HUK。而纤溶酶、尿激酶、凝血酶和胰凝乳蛋白酶不会激活无活性HUK。无活性HUK和HUK的分子量估计分别为4.8×10⁴和4.5×10⁴。胰蛋白酶作用于无活性HUK产生的活性HUK（HUK''）的分子量与HUK几乎相同。在免疫电泳和聚丙烯酰胺凝胶圆盘电泳上，无活性HUK的迁移率略慢于HUK。另一方面，HUK''的电泳迁移率与HUK几乎相同。这两种活性HUK在水解H - Pro - Phe - Arg - MCA的Km值以及各种蛋白酶抑制剂和抗HUK抗体的抑制谱方面没有显著差异。无活性HUK不能通过直接放射免疫测定法（RIA）测量，但胰蛋白酶作用产生的HUK''可以通过RIA测量。
