人红细胞胰岛素受体酪氨酸蛋白激酶活性的测定  

The Measurement of Insulin Receptor Tyrosine Kinase Activity from Human Erythrocytes

作　　者：朱宁[1] 田恩江[2] 王嘉仪[1] 汤新之[1] 

机构地区：[1]天津医科大学生化教研室,300070 [2]天津市内分泌研究所

出　　处：《天津医药》2000年第2期93-95,共3页Tianjin Medical Journal

基　　金：天津市教委科学基金

摘　　要：目的:建立红细胞胰岛素受体酪氨酸蛋白激酶(IRTK)的活性测定法。方法:应用IRTK催化的聚(谷,酪)4:1的磷酸化产物作为抗原,抗磷酸酪氨酸抗体作为第一抗体,羊抗兔lgG-HRP作为第二抗体,建立酶联免疫吸附测定法。结果:批内变异系数为7.8%,批间变异系数为8.6%。20例健康人的红细胞IRTK活性值为(0.184±0.026)nmolPY/(min.mg受体蛋白),36例2型糖尿病患者中有5例活性低于正常范围。胰岛素在10^(-6)mol/L时对IRTK的激活达到最大限度,Mn^(2+)是IRTK的激活剂。结论:此法灵敏、定量、准确、简便,是深入研究糖尿病及其它胰岛素抵抗综合征病因及治疗的一个极有用的检测方法。Objective: To establish a method of detecting insulin receptor tyrosine kinase (IRTK) activity from erythrocytes. Methods:Poly (Glu,Tyr) 4:1 was phosphorylated by catalysis of IRTK. The ELISA system consisted of phosphorylated poly (Glu, Tyr) 4:1 as antigen, antiphosphotyrosine antibody as first antibody and sheep antirabbit lgG-HRP as second anti-body . Results: CV (coefficient of variation) within the batch was 7.8%. CV between the batches was 8.6%. The value of IRTK activity in 20 normal individuals was (0.184±0.026) nmol PY/(min. mg receptor protein). The activity values from 5 of 36 NIDDM patients were below the normal extent. In vitro, IRTK activity reached climax at 10-6 mol/L insulin. Mn2+ was an activator of IRTK. Conclusion:The method is sensitive,quantitative, exact and simple. It is useful for researching the cause and care of insulin resistance.

关 键 词：受体 胰岛素 酪氨酸蛋白激酶 ELISA 糖尿病 

分 类 号：R587.1[医药卫生—内分泌]