人工半合成槲皮素水溶性衍生物对重组人磷脂酰肌醇3—激酶p110β催化亚基的影响

目的:研究人工半合成槲皮素水溶性衍生物—槲皮素-7-硫酸酯钠盐(SQMS)和槲皮素-7,4′-二硫酸酯二钠(SQDS)对重组人磷脂酰肌醇3-激酶(PI3-K)p110β催化亚基的影响.方法:利用基因工程的方法获得PI3-K p110β催化亚基.用磷脂酰肌醇-4,5-二磷酸,[γ-~(32)P]ATP与重组PI3-K p110β催化亚基起保温的方法测定PI3-K的活性;~(32)P标记的磷脂用氯仿和甲醇抽提、板薄层层析和放射自显影来分析.结果:Wortmannin是PI3-K特异的抑制剂,Wortmannin(2.5-20nmol/L)对重组PI3-K p110β亚基有抑制作用;SQMS和SQDS(2.5-20μmol/L)对重组人PI3-K p110β催化亚基有抑制作用.结论:人工半合成槲皮素水溶性衍生物是一种类型的PI3-K抑制剂.重组人PI3-K p110β催化亚基可作为一种较为简便地筛选和开发有效的PI3-K抑制剂的分子靶点.

Effect of semi-synthesized quercetin water-soluble derivatives on recombinant human phosphatidylinositol 3-kinase p110beta catalytic subunit

AIM: To study the effect of semi-synthesized quercetin water-soluble derivatives sodium quercetin-7-sulfate (SQMS) and disodium quercetin-7,4 -disulfate (SQDS) on recombinant human phosphatidylinositol 3-kinase (PI3-K) p110 beta catalytic subunit. METHODS: Recombinant human PI3-K p110 beta catalytic subunit was expressed by gene engineering. PI3 -K was assayed by incubating recombinant PI3-K p110beta with phosphatidylinositol-4,5-bisphosphate and [gamma-32P]ATP; the [32 P]-radiolabeled lipids were extracted with chloroform and methanol, assessed by thin layer chromatography and visualized by autoradiography. RESULTS: Wortmannin, a specific inhibitor o f PI3-K, showed inhibition on recombinant PI3-K p110beta catalytic subunit in a concentration-dependent manner (2.5 - 20 nmol/L); SQMS and SQD S showed inhibition on recombinant PI3-K p110beta catalytic subunit in a concentration-dependent manner (2.5 - 20 micromol/L). CONCLUSION: Semi-synthesized quercetin water-soluble derivatives were a type of inhibitors of PI3-K. The recombinant PI3-K p110beta catalytic subunit might be used as a molecular target for simpler filtrating and development of more effective inhibitors of PI3-K.