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Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging

Insulin stimulation of glucose uptake is achieved by redistribution of insulin-responsive glucose transporters, GLUT4, from intracellular storage compartment(s) to the plasma membrane in adipocytes and muscle cells. Although GLUT4 translocation has been investigated using various approaches, GLUT4 t...

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Autores principales: Fujita, Hideaki, Hatakeyama, Hiroyasu, Watanabe, Tomonobu M., Sato, Masaaki, Higuchi, Hideo, Kanzaki, Makoto
Formato: Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2912357/
https://www.ncbi.nlm.nih.gov/pubmed/20519436
http://dx.doi.org/10.1091/mbc.E10-01-0029
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author Fujita, Hideaki
Hatakeyama, Hiroyasu
Watanabe, Tomonobu M.
Sato, Masaaki
Higuchi, Hideo
Kanzaki, Makoto
author_facet Fujita, Hideaki
Hatakeyama, Hiroyasu
Watanabe, Tomonobu M.
Sato, Masaaki
Higuchi, Hideo
Kanzaki, Makoto
author_sort Fujita, Hideaki
collection PubMed
description Insulin stimulation of glucose uptake is achieved by redistribution of insulin-responsive glucose transporters, GLUT4, from intracellular storage compartment(s) to the plasma membrane in adipocytes and muscle cells. Although GLUT4 translocation has been investigated using various approaches, GLUT4 trafficking properties within the cell are largely unknown. Our novel method allows direct analysis of intracellular GLUT4 dynamics at the single molecule level by using Quantum dot technology, quantitatively establishing the behavioral nature of GLUT4. Our data demonstrate the predominant mechanism for intracellular GLUT4 sequestration in the basal state to be “static retention” in fully differentiated 3T3L1 adipocytes. We also directly defined three distinct insulin-stimulated GLUT4 trafficking processes: 1) release from the putative GLUT4 anchoring system in storage compartment(s), 2) the speed at which transport GLUT4-containing vesicles move, and 3) the tethering/docking steps at the plasma membrane. Intriguingly, insulin-induced GLUT4 liberation from its static state appeared to be abolished by either pretreatment with an inhibitor of phosphatidylinositol 3-kinase or overexpression of a dominant-interfering AS160 mutant (AS160/T642A). In addition, our novel approach revealed the possibility that, in certain insulin-resistant states, derangements in GLUT4 behavior can impair insulin-responsive GLUT4 translocation.
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spelling pubmed-29123572010-10-16 Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging Fujita, Hideaki Hatakeyama, Hiroyasu Watanabe, Tomonobu M. Sato, Masaaki Higuchi, Hideo Kanzaki, Makoto Mol Biol Cell Articles Insulin stimulation of glucose uptake is achieved by redistribution of insulin-responsive glucose transporters, GLUT4, from intracellular storage compartment(s) to the plasma membrane in adipocytes and muscle cells. Although GLUT4 translocation has been investigated using various approaches, GLUT4 trafficking properties within the cell are largely unknown. Our novel method allows direct analysis of intracellular GLUT4 dynamics at the single molecule level by using Quantum dot technology, quantitatively establishing the behavioral nature of GLUT4. Our data demonstrate the predominant mechanism for intracellular GLUT4 sequestration in the basal state to be “static retention” in fully differentiated 3T3L1 adipocytes. We also directly defined three distinct insulin-stimulated GLUT4 trafficking processes: 1) release from the putative GLUT4 anchoring system in storage compartment(s), 2) the speed at which transport GLUT4-containing vesicles move, and 3) the tethering/docking steps at the plasma membrane. Intriguingly, insulin-induced GLUT4 liberation from its static state appeared to be abolished by either pretreatment with an inhibitor of phosphatidylinositol 3-kinase or overexpression of a dominant-interfering AS160 mutant (AS160/T642A). In addition, our novel approach revealed the possibility that, in certain insulin-resistant states, derangements in GLUT4 behavior can impair insulin-responsive GLUT4 translocation. The American Society for Cell Biology 2010-08-01 /pmc/articles/PMC2912357/ /pubmed/20519436 http://dx.doi.org/10.1091/mbc.E10-01-0029 Text en © 2010 by The American Society for Cell Biology
spellingShingle Articles
Fujita, Hideaki
Hatakeyama, Hiroyasu
Watanabe, Tomonobu M.
Sato, Masaaki
Higuchi, Hideo
Kanzaki, Makoto
Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging
title Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging
title_full Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging
title_fullStr Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging
title_full_unstemmed Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging
title_short Identification of Three Distinct Functional Sites of Insulin-mediated GLUT4 Trafficking in Adipocytes Using Quantitative Single Molecule Imaging
title_sort identification of three distinct functional sites of insulin-mediated glut4 trafficking in adipocytes using quantitative single molecule imaging
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2912357/
https://www.ncbi.nlm.nih.gov/pubmed/20519436
http://dx.doi.org/10.1091/mbc.E10-01-0029
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