量子点标记活细胞内GLUT4 蛋白的研究

目的:研究量子点标记活细胞内GLUT4蛋白的方法,用于长时程观察活细胞内GLUT4的转运过程。方法:使用在GLUT4蛋白膜外区构建了myc位点的L6-GLUT4myc细胞系,用胰岛素刺激L6细胞内的GLUT4myc转运到细胞膜上,通过抗体抗原反应先后将一抗9E10和偶联二抗IgG的量子点与特异性位点结合。结果:通过量子点标记固定细胞内GLUT4的实验,证明了标记方法的特异性和灵敏性。量子点能够标记细胞膜表面的GLUT4蛋白并伴随GLUT4的胞吞进入细胞。适当调整实验温度,用量子点标记细胞膜上的GLUT4并且在实验过程结束后将标记了量子点的GLUT4保持在细胞膜表面,能够观察活细胞内GLUT4蛋白内化和胞内循环的过程。结论:发展了量子点标记活细胞内GLUT4的方法,为进一步研究活细胞内GLUT4的转运过程打下了基础。

Research of Labeling GLUT4 in Live Cells Using QDs

Objective: To investigate a strategy of labeling GLUT4 in live cells using QDs for long-term observing GLUT4 translocation in live cells.Methods: L6-GLUT4myc cell line with a myc epitope in the first exofacial loop was used.GLUT4myc was translocated to the membrane after stimulated by insulin,and then labeled by 9E10 and Qdot-IgG.Results: The specificity and sensitivity were proved by labeling GLTU4 in fixed cells with QDs.QDs can be bound and internalized with GLUT4 on the membrane of live cells.The complex of GLUT4 and QDs can be retained on the membrane of live cells by control the cells in a lower temperature,and the internalization and circulation of GLUT4 can be observed.Conclusion: A method of labeling GLUT4 in live cells was developed using QDs for studying translocation of GLUT4 in live cells.