摘要
目的介绍一种组织细胞膜胰岛素受体酪氨酸蛋白激酶(TPK)活力测定方法。方法密度梯度离心分离小鼠肝和骨骼肌细胞膜成分,不进行膜胰岛素受体纯化,用单位膜蛋白胰岛素刺激前后的膜TPK活力改变值,作为评估组织细胞膜胰岛素受体TPK活力的定量指标,并与传统方法的单位膜蛋白麦胚凝集素(WGA)亲和层析纯化后的胰岛素受体胰岛素刺激前后的TPK活力改变值进行比较。结果每mg膜蛋白基础(无胰岛素刺激)TPK活力明显高于每mg膜蛋白纯化后的胰岛素受体TPK活力(P<0.01);每mg膜蛋白经胰岛素刺激的TPK活力改变值与每mg膜蛋白纯化后的胰岛素受体TPK活力改变值无显著差别(P>0.05)。结论组织细胞膜上存在TPK,用胰岛素刺激的组织细胞膜TPK活力改变值,可作为组织细胞膜胰岛素受体特异TPK活力。本方法不需要纯化组织细胞膜胰岛素受体,可作为评价组织细胞膜胰岛素受体功能的实用科研技术。
Objective To introduce a improved method to determine the activity of TPK of membrane insulin receptor in liver and muscle. Methods The membrane of tissue was separated by density gradient centrifugation. The changes of activity of TPK in membrane without purification of insulin receptor. The levels of TPK activities pre-or post-stimulation of the membrane by insulin in liver and muscle represented the activities of TPK of the membrane insulin receptor. Our datas was compared with that of the insulin receptor purified method by WGA-affinity chromatography. Results The activity of TPK of membrane was higher than that of purified insulin receptor;the changes of the activity of TPK of membrane was similar to that of purified insulin receptor. Conclusion The tissue membrane have the activity of TPK, the changes of TPK of membrance stimulated by insulin can be used to determine the activity of TPK of membrane insulin recptor in liver and muscle.
出处
《江西医学院学报》
CAS
2005年第6期57-59,73,共4页
Acta Academiae Medicinae Jiangxi
