Shibata K, Kajihara J, Kato K, Hirano K
Development and Research Laboratories, JCR Pharmaceuticals Co., Ltd., Kobe, Japan.
Biochim Biophys Acta. 1997 Oct 20;1336(3):425-33. doi: 10.1016/s0304-4165(97)00055-x.
Prostate specific antigen (uPSA) was purified to homogeneity from human urine using SuperQ-Toyopearl, Sulfate-Cellulofine, Phenyl-Toyopearl, CM-Sepharose, anti-urokinase IgG Sepharose and Sephadex G-100. The purified uPSA gave a major band at 32.9 kDa on SDS-PAGE under the reduced condition. However, it shows multiple bands on native PAGE. Substrate specificity of purified uPSA is identical with that of PSA from human seminal plasma and uPSA shows the kallikrein and chymotrypsin-like activities. On the analysis of N-terminal amino acid, two amino acid residues at N-terminal position of uPSA were detected and other amino acid sequence of uPSA was identical with that of sPSA. In addition, we isolated the multiple components of uPSA using anion-exchange chromatography. They were almost the same in amino acid composition and N-terminal amino acid sequences and showed differences in lectin-blotting pattern.
使用SuperQ - Toyopearl、硫酸纤维素、苯基 - Toyopearl、CM - 琼脂糖、抗尿激酶IgG琼脂糖和葡聚糖凝胶G - 100从人尿液中纯化前列腺特异性抗原（uPSA）至均一。纯化后的uPSA在还原条件下的SDS - PAGE上呈现一条32.9 kDa的主要条带。然而，它在非变性PAGE上显示多条条带。纯化后的uPSA的底物特异性与来自人精浆的PSA相同，且uPSA具有激肽释放酶和类胰凝乳蛋白酶活性。在对N端氨基酸进行分析时，检测到uPSA N端位置的两个氨基酸残基，uPSA的其他氨基酸序列与sPSA相同。此外，我们使用阴离子交换色谱法分离了uPSA的多种成分。它们在氨基酸组成和N端氨基酸序列上几乎相同，并且在凝集素印迹模式上存在差异。
