C5位上羟基对染料木黄酮抑制重组人蛋白激酶CK2活性的影响

Effect of 5 hydroxyl group of genistein on recombinant human protein kinase CK2 holoenzyme activity

目的观察染料木黄酮C5位上的羟基对其抑制重组人蛋白激酶CK2全酶活性的影响。方法将利用基因工程技术获得的重组人CK2α’及β亚基在体外等摩尔混合构成CK2全酶。通过测定药物作用后转移到CK2底物上的[γ-32P]ATP的32P的放射性活度,探讨染料木黄酮和大豆甙元对重组人CK2全酶活性的抑制作用,并采用半数效量概率单位法计算其IC50值。结果染料木黄酮和大豆甙元能明显抑制重组人CK2全酶活性,其IC50值分别是27.95μmol/L和2.38μmol/L,作用效果接近或者强于目前已知的CK2抑制剂N-(2-氨乙基)-5-氯萘-1-硫胺(A3)。结构效应分析表明:C5位上的羟基可减弱染料木黄酮对重组人CK2全酶的抑制作用。结论染料木黄酮和大豆甙元是两种有效的重组人CK2全酶的抑制剂。

Objective To observed the effects of 5 hydroxyl group of genistein on recombinant human protein kinase CK2 holoenzyme activity. Methods Recombinant human protein kinase CK2α＇ and β subunits which were abtained by genetic engineering teenology were mixed into reconstitute CK2 holoenzyme, the effects of 5 hydroxyl group of genistein on recombinant human protein kinase CK2 holoenzyme activity was study by detecting incorporation of 32 p of [γ-^32 P]ATP. Results Genistein and daidzein were shown to inhibit obviously recombinant CK2 holoenzyme activity in a concentration-dependent manner with IC50 values of 27.95,2.38 μmol/L, respectively, Which display a close effect or more potent than previously known inhibitor N-（2-aminoethyl）-5-chloronaphthalene-l-sulfonamide （A3）. Structure-activity study indicated that hydroxyl group at position 5 is detrimental per se for the inhibitory effects of genistein on CK2. Conclusion Genistein and daidzein were two effective inhibitors of recombinant human protein kinase CK2 in vitro.