Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking

Mark Larance, Georg Ramm, Jacqueline Stöckli, Ellen M. van Dam, Stephanie Winata, Valerie Wasinger, Fiona Simpson, Michael Graham, Jagath R. Junutula, Michael Guilhaus, David E. James (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    279 Citations (Scopus)

    Abstract

    Insulin stimulates the translocation of the glucose transporter GLUT4 from intracellular vesicles to the plasma membrane. In the present study we have conducted a comprehensive proteomic analysis of affinity-purified GLUT4 vesicles from 3T3-L1 adipocytes to discover potential regulators of GLUT4 trafficking. In addition to previously identified components of GLUT4 storage vesicles including the insulin-regulated aminopeptidase insulin-regulated aminopeptidase and the vesicle soluble N-ethylmaleimide factor attachment protein (v-SNARE) VAMP2, we have identified three new Rab proteins, Rab10, Rab11, and Rab14, on GLUT4 vesicles. We have also found that the putative Rab GTPase-activating protein AS160 (Akt substrate of 160 kDa) is associated with GLUT4 vesicles in the basal state and dissociates in response to insulin. This association is likely to be mediated by the cytosolic tail of insulin-regulated aminopeptidase, which interacted both in vitro and in vivo with AS160. Consistent with an inhibitory role of AS160 in the basal state, reduced expression of AS160 in adipocytes using short hairpin RNA increased plasma membrane levels of GLUT4 in an insulin-independent manner. These findings support an important role for AS160 in the insulin regulated trafficking of GLUT4.
    Original languageEnglish
    Pages (from-to)37803-37813
    Number of pages11
    JournalJournal of Biological Chemistry
    Volume280
    Issue number45
    DOIs
    Publication statusPublished - 11 Nov 2005

    Fingerprint

    rab GTP-Binding Proteins
    GTPase-Activating Proteins
    Insulin
    Substrates
    Aminopeptidases
    Cell membranes
    Adipocytes
    Vesicle-Associated Membrane Protein 2
    Cell Membrane
    SNARE Proteins
    Ethylmaleimide
    Facilitative Glucose Transport Proteins
    Proteomics
    Small Interfering RNA
    Proteins

    Keywords

    • Secretory Vesicles
    • Animals
    • Cystinyl Aminopeptidase
    • GTPase-Activating Proteins
    • Mice
    • 3T3-L1 Cells
    • Aminopeptidases
    • Glucose Transporter Type 4
    • Insulin
    • Fibroblasts
    • Gene Expression Profiling
    • Adipocytes
    • Gene Expression Regulation
    • RNA Interference
    • Cell Line
    • Protein Transport
    • Cricetinae

    Cite this

    Larance, M., Ramm, G., Stöckli, J., van Dam, E. M., Winata, S., Wasinger, V., ... James, D. E. (2005). Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking. Journal of Biological Chemistry, 280(45), 37803-37813. https://doi.org/10.1074/jbc.M503897200
    Larance, Mark ; Ramm, Georg ; Stöckli, Jacqueline ; van Dam, Ellen M. ; Winata, Stephanie ; Wasinger, Valerie ; Simpson, Fiona ; Graham, Michael ; Junutula, Jagath R. ; Guilhaus, Michael ; James, David E. / Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 45. pp. 37803-37813.
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    abstract = "Insulin stimulates the translocation of the glucose transporter GLUT4 from intracellular vesicles to the plasma membrane. In the present study we have conducted a comprehensive proteomic analysis of affinity-purified GLUT4 vesicles from 3T3-L1 adipocytes to discover potential regulators of GLUT4 trafficking. In addition to previously identified components of GLUT4 storage vesicles including the insulin-regulated aminopeptidase insulin-regulated aminopeptidase and the vesicle soluble N-ethylmaleimide factor attachment protein (v-SNARE) VAMP2, we have identified three new Rab proteins, Rab10, Rab11, and Rab14, on GLUT4 vesicles. We have also found that the putative Rab GTPase-activating protein AS160 (Akt substrate of 160 kDa) is associated with GLUT4 vesicles in the basal state and dissociates in response to insulin. This association is likely to be mediated by the cytosolic tail of insulin-regulated aminopeptidase, which interacted both in vitro and in vivo with AS160. Consistent with an inhibitory role of AS160 in the basal state, reduced expression of AS160 in adipocytes using short hairpin RNA increased plasma membrane levels of GLUT4 in an insulin-independent manner. These findings support an important role for AS160 in the insulin regulated trafficking of GLUT4.",
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    Larance, M, Ramm, G, Stöckli, J, van Dam, EM, Winata, S, Wasinger, V, Simpson, F, Graham, M, Junutula, JR, Guilhaus, M & James, DE 2005, 'Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking', Journal of Biological Chemistry, vol. 280, no. 45, pp. 37803-37813. https://doi.org/10.1074/jbc.M503897200

    Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking. / Larance, Mark; Ramm, Georg; Stöckli, Jacqueline; van Dam, Ellen M.; Winata, Stephanie; Wasinger, Valerie; Simpson, Fiona; Graham, Michael; Junutula, Jagath R.; Guilhaus, Michael; James, David E. (Lead / Corresponding author).

    In: Journal of Biological Chemistry, Vol. 280, No. 45, 11.11.2005, p. 37803-37813.

    Research output: Contribution to journalArticle

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    T1 - Characterization of the role of the Rab GTPase-activating protein AS160 in insulin-regulated GLUT4 trafficking

    AU - Larance, Mark

    AU - Ramm, Georg

    AU - Stöckli, Jacqueline

    AU - van Dam, Ellen M.

    AU - Winata, Stephanie

    AU - Wasinger, Valerie

    AU - Simpson, Fiona

    AU - Graham, Michael

    AU - Junutula, Jagath R.

    AU - Guilhaus, Michael

    AU - James, David E.

    PY - 2005/11/11

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    N2 - Insulin stimulates the translocation of the glucose transporter GLUT4 from intracellular vesicles to the plasma membrane. In the present study we have conducted a comprehensive proteomic analysis of affinity-purified GLUT4 vesicles from 3T3-L1 adipocytes to discover potential regulators of GLUT4 trafficking. In addition to previously identified components of GLUT4 storage vesicles including the insulin-regulated aminopeptidase insulin-regulated aminopeptidase and the vesicle soluble N-ethylmaleimide factor attachment protein (v-SNARE) VAMP2, we have identified three new Rab proteins, Rab10, Rab11, and Rab14, on GLUT4 vesicles. We have also found that the putative Rab GTPase-activating protein AS160 (Akt substrate of 160 kDa) is associated with GLUT4 vesicles in the basal state and dissociates in response to insulin. This association is likely to be mediated by the cytosolic tail of insulin-regulated aminopeptidase, which interacted both in vitro and in vivo with AS160. Consistent with an inhibitory role of AS160 in the basal state, reduced expression of AS160 in adipocytes using short hairpin RNA increased plasma membrane levels of GLUT4 in an insulin-independent manner. These findings support an important role for AS160 in the insulin regulated trafficking of GLUT4.

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    KW - Secretory Vesicles

    KW - Animals

    KW - Cystinyl Aminopeptidase

    KW - GTPase-Activating Proteins

    KW - Mice

    KW - 3T3-L1 Cells

    KW - Aminopeptidases

    KW - Glucose Transporter Type 4

    KW - Insulin

    KW - Fibroblasts

    KW - Gene Expression Profiling

    KW - Adipocytes

    KW - Gene Expression Regulation

    KW - RNA Interference

    KW - Cell Line

    KW - Protein Transport

    KW - Cricetinae

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