Tyrphostin AG555对重组人肌醇磷脂3-激酶p110β催化亚基活性的影响

Effect of tyrphostin AG555 on the catalytic subunit activity of the recombinant human phosphoinositide 3-kinase p110β

目的:研究tyrphostin AG555对重组人肌醇磷脂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-32P]ATP与重组 PI3-K p110β催化亚基一起保温的方法测定PI3-K的活性; 用氯仿和甲醇抽提32P标记的磷脂,并以硅胶板薄层层析和放射自显影来进行分析.结果:AG555 (2.5～20 μmol/L)对重组人 PI3-K p110β催化亚基有抑制作用.结论:tyrphostin AG555是一种重组人PI3-Kp110β抑制剂.重组人PI3-K p110β催化亚基可作为一种较为简便的筛选有效的PI3-K抑制剂的反应物.

Objective： To observe the effect of tyrphostin AG555 on the catalytic subunit activity of the recombinant human phosphoinositide 3-kinase （PI3-K） p110β. Methods： The recombinant human PI3-K p100β catalytic subunit was obtained by genetic engineering： the activity of PI3-K was detected by incubating the recombinant PI3-K p110β with phosphatidylinositol-4,5-bisphosphate and [У-^32P]ATP at stable temperature; the resulting ^32P-radiolabeled lipids were extracted with chloroform and methanol, then assessed by the thin layer chromatography and autoradiography. Results： AG555 inhibited the catalytic subunit activity of the recombinant PI3-K p110β in a concentration dependent manner （2.5～20μmol/L）. Conclusion; AG555 might be an potential inhibitor of PI3-K p110β which could be used as a molecular target for simply screening potential inhibitors of PI3-K.