Direct Quantification of Fusion Rate Reveals a Distal Role for AS160 in Insulin-stimulated Fusion of GLUT4 Storage Vesicles.

Direct Quantification of Fusion Rate Reveals a Distal Role for AS160 in Insulin-stimulated Fusion of GLUT4 Storage Vesicles. Research Abstract Details 

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Direct Quantification of Fusion Rate Reveals a Distal Role for AS160 in Insulin-stimulated Fusion of GLUT4 Storage Vesicles. Abstract Text:

Li Jiang,Junmei Fan,Li Bai,Yan Wang,Yu Chen,Lu Yang,Liangyi Chen,Tao Xu,

Insulin-stimulated GLUT4 translocation to the plasma membrane constitutes a key process for blood glucose control. However, convenient and robust assays to monitor this dynamic process in real time are lacking, which hinders current progress toward elucidation of the underlying molecular events as well as screens for drugs targeting this particular pathway. Here, we have developed a novel dual colored probe to monitor the translocation process of GLUT4 based on dual color fluorescence measurement. We demonstrate that this probe is more than an order of magnitude more sensitive than the current technology for detecting fusion events from single GLUT4 storage vesicles (GSVs). A small fraction of fusion events were found to be of the "kiss-and-run" type. For the first time, we show that insulin stimulation evokes a approximately 40-fold increase in the fusion of GSVs in 3T3-L1 adipocytes, compared with basal conditions. The probe can also be used to monitor the prefusion behavior of GSVs. By quantifying both the docking and fusion rates simultaneously, we demonstrate a proportional inhibition in both docking and fusion of GSVs by a dominant negative mutant of AS160, indicating a role for AS160 in the docking of GSVs but not in the regulation of GSV fusion after docking.

Direct Quantification of Fusion Rate Reveals a Distal Role for AS160 in Insulin-stimulated Fusion of GLUT4 Storage Vesicles. Publishing Authors By Initials

L Jiang,J Fan,L Bai,Y Wang,Y Chen,L Yang,L Chen,T Xu,

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Direct Quantification of Fusion Rate Reveals a Distal Role for AS160 in Insulin-stimulated Fusion of GLUT4 Storage Vesicles. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: The Journal of biological chemistry

VOLUME: 283

Page Numbers: 8508-16

Journal Abbreviation: J. Biol. Chem.

ISSN: 0021-9258

DAY: 6

MONTH: 12

YEAR: 2007

Direct Quantification of Fusion Rate Reveals a Distal Role for AS160 in Insulin-stimulated Fusion of GLUT4 Storage Vesicles. Information

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LANGUAGE: eng

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AFFILIATION: Joint Laboratory of Institute of Biophysics & Huazhong University of Science and Technology, National Laboratory of Biomacromolecules, Chinese Academy of Sciences, Beijing 100101, China.

Country: United States

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MEDLINETA: J Biol Chem

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