Fujimoto Y, Suzuki C, Watanabe Y, Matsuda Y, Akihama S
Department of Clinical Biochemistry, Hokkaido Institute of Pharmaceutical Sciences, Japan.
Biochem Med Metab Biol. 1990 Dec;44(3):218-27. doi: 10.1016/0885-4505(90)90064-8.
A tissue kallikrein was purified over 1500-fold from the postmicrosomal supernatant of human submaxillary glands. The purified enzyme gave a single band, corresponding to an apparent molecular weight of 42,000 on SDS-polyacrylamide gel electrophoresis. This enzyme cross-reacted with the anti-human urinary kallikrein antiserum. The purified enzyme was characterized in comparison with the purest human urinary kallikrein preparation. Both enzymes hydrolyzed the synthetic substrate, Ac-Phe-Arg-OMe, most effectively. Aprotinin, TLCK, and PMSF suppressed the enzyme activities, while SBTI, LBTI, and alpha 1-antitrypsin had no effect at all. The purified enzyme generated kinin from the natural substrate, kininogen. It was concluded therefore that the purified enzyme is a typical tissue kallikrein.
从人下颌下腺微粒体后上清液中纯化出一种组织激肽释放酶,纯化倍数超过1500倍。纯化后的酶在SDS-聚丙烯酰胺凝胶电泳上呈现单一条带,对应表观分子量为42,000。该酶与抗人尿激肽释放酶抗血清发生交叉反应。将纯化后的酶与最纯的人尿激肽释放酶制剂进行比较表征。两种酶对合成底物Ac-Phe-Arg-OMe的水解效果最佳。抑肽酶、TLCK和PMSF可抑制酶活性,而SBTI、LBTI和α1-抗胰蛋白酶则完全没有作用。纯化后的酶可从天然底物激肽原生成激肽。因此得出结论,纯化后的酶是一种典型的组织激肽释放酶。