毛细管区带电泳测定D1蛋白酶的活性及其抑制剂先导化合物的筛选  

Determination of D1 Protease Activity and Screening for Lead Compounds of Inhibitor by Capillary Zone Electrophoresis

作　　者：陈腊月[1] 李伟国[2] 黄雪英[2] 刘中来[1] 刘艳丽[1] 冯璐 廖宏珊 熊国梅[1] 祁超[1] 

机构地区：[1]华中师范大学生命科学学院遗传调控与整合生物学湖北省重点实验室 [2]华中师范大学化学学院农药与化学生物学教育部重点实验室,武汉430079

出　　处：《分析化学》2011年第11期1758-1761,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.20702017,30670429);湖北省自然科学基金(Nos.2007ABA141,2010CDB01206,2009CDB234)资助

摘　　要：建立了毛细管区带电泳技术快速测定D1蛋白酶活性及其抑制剂先导化合物的筛选方法。实验选用未涂层熔融石英毛细管(43 cm 5mm)和磷酸盐缓冲溶液(50 mmol/L, pH 3.0)作为分离介质,运行电压18 kV,测定了D1蛋白酶的活性并对部分抑制剂先导化合物进行了筛选。结果表明, ITP26和ITP21两种异噁唑噻唑哌啶类先导化合物对蛋白酶活性具有抑制作用,抑制率分别为26%和13%。Capillary zone electrophoresis （CZE） wi＊h UV detection has been used for the determination of D1 protease activity and screening for lead compounds of inhibitor. Separation conditions of peptides were optimized as phosphate buffer （50 mmol/L, pH 3.0） at 18 kV running voltage, with capillary length of 43 cm ×75 μm （id）. Hydrolysis activity of D1 protease and screening for lead compounds of inhibitor was performed by CZE. The results indicated that two lead compounds ITP 21 and ITP 26 showed inhibitory effects of 26% and 13%, respectively, while other compounds had no inhibitory effects on D1 protease activity. This method is simple and convenient, suitable for the screening study on protease＇s inhibitors.

关 键 词：D1蛋白酶 毛细管区带电泳 活性 先导化合物 

分 类 号：Q55[生物学—生物化学] O657.8[理学—分析化学]