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Dynamics of insulin‐stimulated translocation of GLUT4 in single living cells visualised using green fluorescent protein
Author(s) -
Dobson Stephen P.,
Livingstone Callum,
Gould Gwyn W.,
Tavaré Jeremy M.
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00879-4
Subject(s) - glut4 , green fluorescent protein , fluorescence , chromosomal translocation , dynamics (music) , chemistry , insulin , protein dynamics , microbiology and biotechnology , biophysics , biochemistry , biology , protein structure , endocrinology , gene , physics , optics , acoustics
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.