毛细管电泳测定鸡肝二氢叶酸还原酶活力及其抑制剂的抑制率  

Determination of Dihydrofolate Reductase Activity and the Inhibitory Rate of Its Inhibitors by Capillary Electrophoresis

作　　者：栗娜[1] 张馨月[1] 朱若华[1] 

机构地区：[1]首都师范大学化学系,北京100037

出　　处：《分析科学学报》2008年第6期631-635,共5页Journal of Analytical Science

基　　金：北京市教委资助项目(No.KM200610028007)

摘　　要：本文采用毛细管电泳法对亲和层析提取的鸡肝二氢叶酸还原酶(DHFR)的反应体系进行了研究,根据体系中反应物还原辅酶Ⅱ(NADPH)和反应产物氧化型辅酶Ⅱ(NADP)的浓度变化,测得鸡肝二氢叶酸还原酶的米氏常数Km=1.35×10-5mol/L。将该体系用于鸡肝二氢叶酸还原酶抑制剂(DHFRIs)抑制率的测定,测得氨甲喋呤的半数抑制浓度(Ic50)为7.46×10-8mol/L。Capillary electrophoresis （CE） has been applied to the study of the dihydrofolate reductase which was purified from chicken liver by affinity chr difference of peak areas between reactants （NADPH）and products（NADP） was us reaction system of omatography. The ed to determine the dynamic parameter Km of DHFR. The value of Km is 1.35×10^-5mol/L. The proposed CE method was used to determine the inhibition rate of methotrexate（MTX） to dihydrofolate reduetase. The Icso of MTX is 7.46×10^-8mol/L.

关 键 词：毛细管电泳 二氢叶酸还原酶 抑制剂 酶活 

分 类 号：O657.8[理学—分析化学]