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

Inhibitory effect and kinetic analysis of 1,8-dihydroxyanthraquinone on recombinant human protein kinase CK2 holoenzyme

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. 90μmol·L-1; 与ATP呈竞争性抑制CK2的活性, 抑制常数Ki值为16. 08μmol·L-1。结论1, 8 -二羟蒽醌是重组人蛋白激酶CK2的抑制剂。

Objective The direct effect and kinetics of 1,8-dihydroxyanthraquinone on holoenzyme were investigated to evaluate the inhibitor of protein kinase CK2. Methods The effects of a series of 1,8-dihydroxyanthraquinone of different concentrations on recombinant human protein kinase CK2 holoenzyme were investigated. The CK2 activity was assayed by detecting incorporation of 32P of [γ-32P] ATP into the substrate in various conditions. Results The characterization and function of the reconstituted holoenzyme were consistent with those of native CK2. 1,8-dihydroxyanthraquinone strongly inhibited the holoenzyme activity of recombinant human protein kinase CK2 (IC (50)=23.90 μmol·L^(-1)). Kinetic studies of 1,8-dihydroxyanthraquinone on CK2 showed that the inhibition was competitive with ATP (K i=16.08 μmol·L^(-1)). Conclusions 1,8-dihydroxyanthraquinone is an effective inhibitor of CK2 holoenzyme.