NDLI: Self-assembly of Glut4 storage vesicles during differentiation of 3T3-L1 adipocytes.

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Self-assembly of Glut4 storage vesicles during differentiation of 3T3-L1 adipocytes.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Shi, Jun Kandror, Konstantin V. Huang, Guanrong
Description	Author Affiliation: Shi J ( Boston University School of Medicine, Boston, Massachusetts 02118, USA.)
Abstract	Glut4 storage vesicles (GSVs) represent translocation-competent vesicular carriers in fat and skeletal muscle cells that deliver Glut4 to the plasma membrane in response to insulin stimulation. GSVs include three major cargo proteins: Glut4, insulin-responsive aminopeptidase (IRAP), and sortilin. Previous work has suggested that the lumenal interaction between Glut4 and sortilin and the cytoplasmic interaction between sortilin and GGA adaptors play an important role in recruitment of Glut4 into the GSVs. However, the mechanism of IRAP targeting to this compartment remains unknown. To address this question, we show that in differentiating adipocytes IRAP enters the GSVs from the 'donor' membranes on day 3 of differentiation. Forced expression of sortilin in undifferentiated cells does not recruit IRAP into the vesicles. However, double expression of sortilin and Glut4 reconstitutes functional GSVs that incorporate endogenous IRAP. To explain this process, we show by a yeast two-hybrid system and chemical cross-linking that the lumenal domain of IRAP can interact with the lumenal loop of Glut4. IRAP without the lumenal domain is faithfully targeted to the donor membranes but has significantly lower insulin responsiveness than full-length IRAP. We suggest that lumenal interactions between Glut4 and IRAP play an important role in the assembly of the GSVs.
ISSN	00219258
e-ISSN	1083351X
Journal	Journal of Biological Chemistry
Issue Number	44
Volume Number	283
Language	English
Publisher	American Society for Biochemistry and Molecular Biology (United States)
Publisher Date	2008-10-31
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Adipocytes Metabolism Cystinyl Aminopeptidase Glucose Transporter Type 4 Physiology 3T3-L1 Cells Adaptor Proteins, Vesicular Transport Animals Cell Differentiation Cross-Linking Reagents Pharmacology Cytosol Insulin Membrane Glycoproteins Mice Models, Biological Nerve Tissue Proteins Protein Conformation Protein Structure, Tertiary Two-Hybrid System Techniques Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Biochemistry Molecular Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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