基于磁分离方法的猪胰岛素受体分离与活性分析  

Separation Based on Magnetic Probes and Activity Identification of Insulin Receptor in Liver of Pig

作　　者：钟王磊 陈维娴 周国华 ZHONG Wang-lei;CHEN Wei-xian;ZHOU Guo-hua(Life Science & Technology School,School of Chemistry and Chemical Engineering,Lingnan Normal University,Zhanjiang 524048,China;Key Laboratory of Clean Energy Materials Chemistry of Guangdong Higher Education Institutes,School of Chemistry and Chemical Engineering,Lingnan Normal University,Zhanjiang 524048,China)

机构地区：[1]岭南师范学院生命科学与技术学院,广东湛江524048 [2]岭南师范学院化学化工学院清洁能源材料化学广东普通高校重点实验室,广东湛江524048

出　　处：《化学试剂》2019年第9期982-986,共5页Chemical Reagents

基　　金：国家自然科学基金资助项目(21605074);广东省自然科学基金资助项目(2014A030310274);稀土资源利用与发光材料研究创新团队项目(2017KCXTD022);岭南师范学院自然科学基金资助项目(ZL1404)

摘　　要：目前,胰岛素受体的分离提取方法为凝集素亲和层析与以胰岛素作为配体的亲和层析。建立磁分离法,采用胰岛素受体亲和磁分离探针提取受体蛋白,对提取条件进行优化,并以免疫印迹法检测胰岛素作用下受体蛋白酪氨酸的自磷酸化反应。结果表明,磁分离探针与粗提取总蛋白孵育16 h是提取受体蛋白的较优条件,所得胰岛素受体保持了其生物活性,能够发生自磷酸化反应。这些结果表明磁分离方法可应用于胰岛素受体的分离纯化,该方法简便、特异性强、稳定、可靠,为胰岛素信号传导、糖尿病病理、药物筛选等研究提供了有效的研究工具。At present,lectin affinity chromatography and insulin affinity chromatography are two main methods of separation and purification of insulin receptor proteins.A magnetic separation probe was designed to isolate and purify insulin receptors and tested the biological activity of the extracted receptor proteins.Receptor protein was extracted by insulin receptor affinity magnetic separation probe,and the concentration of receptor protein extracted under different conditions was compared.The tyrosine auto-phosphorylation of receptor protein with insulin stimulus was detected by western blotting.The results showed that 16 hours of incubation between the magnetic separation probe and the total protein was the best condition for extracting the receptor protein.The tyrosine auto-phosphorylation of insulin receptor showed that the isolated insulin receptor has biological activity and can be used for further study of insulin signal transduction.Therefore,the magnetic separation method can be applied to the separation and purification of insulin receptor.

关 键 词：胰岛素受体 磁分离 提取 信号传导 自磷酸化 

分 类 号：Q5-33[生物学—生物化学]