1,8二羟蒽醌对重组人蛋白激酶CK2全酶的抑制作用及其动力学分析
重组蛋白,,蛋白激酶CK2；重组蛋白；全酶；蒽醌；1,8二羟蒽醌；酶动力学,1材料与方法,2结果,3讨论,参考文献
摘 要 目的 观察1,8二羟蒽醌对重组人蛋白激酶CK2全酶的直接作用及其酶动力学机制，以寻找CK2的抑制剂。方法 以重组人CK2全酶为分子靶点，检测不同浓度1,8二羟蒽醌对CK2活性的影响。CK2活性通过测定转移到CK2底物上的[γ32P]ATP的[32P]放射活度来检测。结果 重组人CK2与天然CK2的性质完全一致。1,8二羟蒽醌对重组人CK2全酶具有较强的抑制作用，半数抑制浓度(IC50)为23.90 μmol·L-1; 与ATP呈竞争性抑制CK2的活性, 抑制常数Ki值为16.08 μmol·L-1。结论 1,8二羟蒽醌是重组人蛋白激酶CK2的抑制剂。关键词 蛋白激酶CK2；重组蛋白；全酶；蒽醌；1,8二羟蒽醌；酶动力学
Inhibitory effect and kinetic analysis of 1,8dihydroxyanthraquinone on recombinant human protein kinase CK2 holoenzyme
LI Chunmei, LIU Xinguang, LIN Xiaocong, CHEN Xiaowen, LIANG Nianci
(1.Department of Biochemistry, Guangdong College of Pharmacy, Guangzhou 510240, China; 2.Institute of Biochemistry & Molecular Biology, Guangdong Medical College, Zhanjiang 524023, PRC; 3.Paediatrics Institute of Shenzhen Childrens Hospital, Shenzhen 518026, China)
Abstract Objective The direct effect and kinetics of 1,8dihydroxyanthraquinone on holoenzyme were investigated to evaluate the inhibitor of protein kinase CK2. Methods The effects of a series of 1,8dihydroxyanthraquinone of different concentrations on recombinant human protein kinase CK2 holoenzyme were investigated. The CK2 activity was assayed by detecting incorporation of 32P of [γ32P] ATP into the substrate in various conditions. Results The characterization and function of the reconstituted holoenzyme were consistent with those of native CK2. 1,8dihydroxyanthraquinone strongly inhibited the holoenzyme activity of recombinant human protein kinase CK2 (IC50=23.90 μmol·L-1). Kinetic studies of 1,8dihydroxyanthraquinone on CK2 showed that the inhibition was competitive with ATP (Ki=16.08 μmol·L-1). Conclusions 1,8dihydroxyanthraquinone is an effective inhibitor of CK2 holoenzyme. ......
您现在查看是摘要页，全文长 9533 字符。