摘要
目的建立一种能够检测不同分子大小(尤其是低分子量)蛋白酶抑制剂的电泳方法。方法根据特异蛋白酶抑制剂抑制靶蛋白酶活性的原理,借助明胶-Tricine-SDS-聚丙烯酰胺凝胶电泳分离样品,胶板经蛋白酶水解后,以考马斯亮蓝染色。结果小分子量的胃蛋白酶抑制剂与较大分子量的BBI均能在该方法中显示出活性带,并且利用该方法检测到山合欢种子中含有一种胰蛋白酶抑制剂。结论该方法能够满足不同分子大小、不同类型蛋白酶抑制剂检测的需要。山合欢胰蛋白酶抑制剂的发现对山合欢进一步的研究和综合开发有重要意义。
Objective To establish an detecting method for protease inhibitors, especially for low - molecular - weight inhibitors. Methods Inhibitor samples were separated on a gelatin - SDS - PAGE in a Tris - Tricine buffer system. After electrophoresis, the gel was incubated with the target proteases to hydrolyze the background gelatin. The inhibitor bands, which were undi- gested by the target proteases, were stained. Results Low - molecular - weight inhibitor （ pepstatin A） and larger inhibitor （ soybean Bowman - Birk inhibitor） were demonstrated by this method and showed clear blue inhibitor bands in the white background. By this method, a trypsin inhibitor was detected from the seeds of Albizzia kalkora （ Roxb. ） Prain. Conclusion This method not only fit for inhibitors with different molecular mass, but fits for inhibitors with various species. Moreover, the trypsin inhibitor found in Albizzia kalkora is important for further research and overall exploration.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2008年第6期1357-1358,共2页
Lishizhen Medicine and Materia Medica Research
基金
西南交通大学科技发展基金项目(No.20060116)
关键词
山合欢
Tricine系统
蛋白酶抑制
电泳
Albizzia kalkora （Roxb.） Prain
Tricinc buffer system
Protease inhibitor
Electrophoresis
