The Amino Terminus of Insulin-responsive Aminopeptidase Causes Glut4 Translocation in 3T3-L1 Adipocytes

The insulin-responsive aminopeptidase (IRAP) is a constituent of the vesicles that contain the insulin-regulated glucose transporter (Glut4). Like Glut4, IRAP translocates to the cell surface in response to insulin. Microinjection into 3T3-L1 adipocytes of a glutathione S-transferase (GST) fusion pr...

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Bibliographic Details
Published in:	The Journal of biological chemistry Vol. 272; no. 37; pp. 23323 - 23327
Main Authors:	Waters, Steven B., D'Auria, Matt, Martin, Stuart S., Nguyen, Chris, Kozma, Lynn M., Luskey, Kenneth L.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 12-09-1997 American Society for Biochemistry and Molecular Biology
Subjects:	3T3 Cells Adipocytes - cytology Adipocytes - drug effects Adipocytes - metabolism ADIPOSE TISSUES AMINOPEPTIDASA AMINOPEPTIDASE Aminopeptidases - genetics Aminopeptidases - pharmacology Androstadienes - pharmacology Animals BINDING PROTEINS Biological Transport Cell Compartmentation Cell Membrane - drug effects CELL MEMBRANES CELLS CELLULE CELULAS CONTROL HORMONAL CONTROLE HORMONAL ESTIMULO Fluorescent Antibody Technique GLUCOSA GLUCOSE GLUCOSE TRANSPORTER Glucose Transporter Type 4 GLUTATHIONE TRANSFERASE HORMONAL CONTROL IMMUNOCYTOCHEMISTRY IMMUNOLOGIE IMMUNOLOGY INMUNOLOGIA INSULIN Insulin - pharmacology INSULINA INSULINE MEMBRANAS CELULARES MEMBRANE CELLULAIRE METABOLISME DES PROTEINES METABOLISMO PROTEICO MICE Monosaccharide Transport Proteins - metabolism Muscle Proteins Peptide Fragments - pharmacology Phosphatidylinositol 3-Kinases Phosphotransferases (Alcohol Group Acceptor) - metabolism PLASMA MEMBRANES PROTEIN METABOLISM PROTEIN TRANSPORT PROTEINAS PROTEINAS AGLUTINANTES PROTEINAS RECOMBINANTES PROTEINE PROTEINE DE LIAISON PROTEINE RECOMBINANTE PROTEINS RATON Recombinant Fusion Proteins - pharmacology RECOMBINANT PROTEINS SOURIS STIMULI STIMULUS SURFACE PROTEINS TEJIDO ADIPOSO TISSU ADIPEUX TRANSFERASAS TRANSFERASE TRANSFERASES Wortmannin
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Summary:	The insulin-responsive aminopeptidase (IRAP) is a constituent of the vesicles that contain the insulin-regulated glucose transporter (Glut4). Like Glut4, IRAP translocates to the cell surface in response to insulin. Microinjection into 3T3-L1 adipocytes of a glutathione S-transferase (GST) fusion protein containing the cytosolic portion of IRAP (GST-IRAP-(1–109)), resulted in translocation of Glut4 to the cell surface. Immunostaining of 3T3-L1 adipocytes for Glut4 showed that the percentage of cells with substantial cell surface Glut4 was 10% in unstimulated cells, 8% following injection of GST, and 27% following injection of GST-IRAP-(1–109). Increased cell surface Glut4 occurred within 5–10 min following injection and was maintained for at least 4 h. A fusion protein containing only 28 amino acids from IRAP (GST-IRAP-(55–82)) was as effective in increasing cell surface Glut4 as stimulation with 100 nm insulin (44% versus43%, respectively). In contrast to insulin-stimulated Glut4 translocation, the redistribution of Glut4 following injection of GST-IRAP-(55–82) was not blocked by wortmannin or co-injection with a SH2 domain from the regulatory subunit of phosphatidylinositol 3-kinase. These data suggest that the amino terminus of IRAP interacts with a retention/sorting protein that also regulates the distribution of Glut4 in insulin-responsive cells.
Bibliography:	S20 1997082018 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0021-9258 1083-351X
DOI:	10.1074/jbc.272.37.23323