Clathrin-dependent and independent endocytosis of glucose transporter 4 (GLUT4) in myoblasts: regulation by mitochondrial uncoupling

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Costin N AntonescuAmira Klip

Abstract

In myocytes and adipocytes, insulin increases glucose transporter 4 (GLUT4) exocytosis by promoting GLUT4 vesicle docking/fusion with the membrane. Less is known about the mechanism and regulation of GLUT4 endocytosis, particularly in myocytes. Here, we show that GLUT4 internalization in L6 myoblasts was inhibited in part by hypertonicity or clathrin heavy chain knockdown and in part by cholesterol depletion. Both strategies had additive effects, abolishing GLUT4 endocytosis. GLUT4 internalization was abrogated by expressing dominant-negative dynamin-2 but unaffected by inhibiting caveolar-dependent endocytosis through syntaxin-6 knockdown or caveolin mutants (which reduced lactosylceramide endocytosis). Insulin did not affect GLUT4 internalization rate or sensitivity to clathrin or cholesterol depletion. In contrast, the mitochondrial uncoupler dinitrophenol (DNP), which like insulin increases surface GLUT4, reduced GLUT4 (but not transferrin) internalization, an effect additive to that of depleting clathrin but not cholesterol. Trout GLUT4 (a natural variant of GLUT4 bearing different endocytic motifs) exogenously expressed in mammalian L6 cells internalized only through the cholesterol-dependent route that also included the ...Continue Reading

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Citations

Mar 17, 2011·Biochemistry·Kevin FoleyAmira Klip
Nov 17, 2009·The Journal of Biological Chemistry·Daniel J FazakerleyAdelle C F Coster
Jan 22, 2011·The Journal of Biological Chemistry·Estifanos N HabtemichaelCynthia Corley Mastick
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May 19, 2009·Applied Physiology, Nutrition, and Metabolism = Physiologie Appliquée, Nutrition Et Métabolisme·Amira Klip
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Mar 16, 2011·Traffic·Alexander F RowlandDavid E James
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Oct 18, 2019·Traffic·Sadia RahmaniCostin N Antonescu
Mar 18, 2010·Current Protocols in Cell Biology·Shuhei IshikuraAmira Klip
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Jun 24, 2008·The Biochemical Journal·Hilal ZaidAmira Klip
Jan 16, 2009·Bioscience Reports·Costin N AntonescuAmira Klip

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