Dynamics of insulin‐stimulated translocation of GLUT4 in single living cells visualised using green fluorescent protein

FEBS Letters

Tavaré, Jeremy M.¹  Gould, Gwyn W.²  Dobson, Stephen P.¹  Livingstone, Callum² 

[1] Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK;Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK

关键词: Glucose transport;    Translocation;    Insulin;    Green fluorescent protein;   

DOI  :  10.1016/0014-5793(96)00879-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Insulin increases glucose uptake by promoting the translocation of the GLUT4 isoform of glucose transporters to the plasma membrane. We have studied this process in living single cells by fusing green fluorescent protein (GFP) to the N-terminus (GFP·GLUT4) or C-terminus (GLUT4·GFP) of GLUT4. Both chimeras were expressed in a perinuclear compartment of CHO cells, and in a vesicular distribution through the cytosol. Insulin promoted an increase in plasma membrane fluorescence as a result of net translocation of the chimeras to the cell surface. GLUT4·GFP, but not GFP·GLUT4, was re-internalised upon the removal of insulin suggesting that a critical internalisation signal sequence exists in the N-terminus of GLUT4. The use of GFP thus allows an analysis of GLUT4 trafficking in single living cells.

【 授权许可】

Unknown