A Role for Phospholipase D in GLUT4 Glucose Transporter Translocation

Based on recent studies showing that phospholipase D (PLD)1 is associated with intracellular membranes and promotes membrane budding from the trans-Golgi, we tested its possible role in the membrane trafficking of GLUT4 glucose transporters. Using immunofluorescence confocal microscopy, expressed My...

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Bibliographic Details
Published in:The Journal of biological chemistry 2000-03, Vol.275 (10), p.7144-7151
Main Authors: Emoto, Masahiro, Klarlund, Jes K., Waters, Steve B., Hu, Vivian, Buxton, Joanne M., Chawla, Anil, Czech, Michael P.
Format: Article
Language:English
Subjects:
3T3 Cells
ADP-Ribosylation Factors - physiology
Animals
Biological Transport
CHO Cells
Cricetinae
Glucose Transporter Type 4
glucose transporters
GLUT4 protein
green fluorescent protein
insulin
Insulin - pharmacology
Isoenzymes - analysis
Mice
Monosaccharide Transport Proteins - metabolism
Muscle Proteins
Phospholipase D - analysis
Phospholipase D - physiology
Tetradecanoylphorbol Acetate - pharmacology
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Summary:Based on recent studies showing that phospholipase D (PLD)1 is associated with intracellular membranes and promotes membrane budding from the trans-Golgi, we tested its possible role in the membrane trafficking of GLUT4 glucose transporters. Using immunofluorescence confocal microscopy, expressed Myc epitope-tagged PLD1 was found to associate with intracellular vesicular structures by a mechanism that requires its N-terminal pleckstrin homology domain. Partial co-localization with expressed GLUT4 fused to green fluorescent protein in both 3T3-L1 adipocytes and Chinese hamster ovary cells was evident. Furthermore, microinjection of purified PLD into cultured adipocytes markedly potentiated the effect of a submaximal concentration of insulin to stimulate GLUT4 translocation to cell surface membranes. Insulin stimulated PLD activity in cells expressing high levels of insulin receptors but no such insulin effect was detected in 3T3-L1 adipocytes. Taken together, these results are consistent with the hypothesis that PLD1 associated with GLUT4-containing membranes acts in a constitutive manner to promote the mechanism of GLUT4 translocation by insulin.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.275.10.7144