GLUT4 translocation by insulin in intact muscle cells: detection by a fast and quantitative assay

FEBS Letters

Ebina, Yousuke¹  Klip, Amira²  Wang, Qinghua²  Khayat, Zayna²  Kishi, Kazuhiro¹ 

[1] Division of Molecular Genetics, Institute for Enzyme Research, The University of Tokushima, Tokushima 770, Japan;Programme in Cell Biology, The Hospital for Sick Children, 555 University Avenue, Toronto, Ont. M5G 1X8, Canada

关键词: Glucose transport;    Cell surface;    Plasma membrane;    Actin cytoskeleton;    Latrunculin B;   

DOI  :  10.1016/S0014-5793(98)00423-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We report a rapid and sensitive colorimetric approach to quantitate the amount of glucose transporters exposed at the surface of intact cells, using L6 muscle cells expressing GLUT4 containing an exofacial myc epitope. Unstimulated cells exposed to the surface 5 fmol GLUT4myc per mg protein. This value increased to 10 fmol/mg protein in response to insulin as 2-deoxyglucose (10 μM) uptake doubled. The results are substantiated by immunofluorescent detection of GLUT4myc in unpermeabilized cells and by subcellular fractionation. We further show that wortmannin and the cytoskeleton disruptors cytochalasin D and latrunculin B completely blocked these insulin effects. The rapid quantitative assay described here could be of high value to study insulin signals and to screen for potential anti-diabetic drugs.

【 授权许可】

Unknown