Insulin regulates Rab3–Noc2 complex dissociation to promote GLUT4 translocation in rat adipocytes

Francoise Koumanov, Vinit J. Pereira, Judith D. Richardson, Samantha L. Sargent, Daniel J. Fazakerley, Geoffrey D. Holman

Research output: Contribution to journalArticle

8 Citations (Scopus)
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Abstract

Aims/hypothesis

The glucose transporter GLUT4 is present mainly in insulin-responsive tissues of fat, heart and skeletal muscle and is translocated from intracellular membrane compartments to the plasma membrane (PM) upon insulin stimulation. The transit of GLUT4 to the PM is known to be dependent on a series of Rab proteins. However, the extent to which the activity of these Rabs is regulated by the action of insulin action is still unknown. We sought to identify insulin-activated Rab proteins and Rab effectors that facilitate GLUT4 translocation.

Methods

We developed a new photoaffinity reagent (Bio-ATB-GTP) that allows GTP-binding proteomes to be explored. Using this approach we screened for insulin-responsive GTP loading of Rabs in primary rat adipocytes.

Results

We identified Rab3B as a new candidate insulin-stimulated G-protein in adipocytes. Using constitutively active and dominant negative mutants and Rab3 knockdown we provide evidence that Rab3 isoforms are key regulators of GLUT4 translocation in adipocytes. Insulin-stimulated Rab3 GTP binding is associated with disruption of the interaction between Rab3 and its negative effector Noc2. Disruption of the Rab3–Noc2 complex leads to displacement of Noc2 from the PM. This relieves the inhibitory effect of Noc2, facilitating GLUT4 translocation.

Conclusions/interpretation

The discovery of the involvement of Rab3 and Noc2 in an insulin-regulated step in GLUT4 translocation suggests that the control of this translocation process is unexpectedly similar to regulated secretion and particularly pancreatic insulin-vesicle release.
Original languageEnglish
Pages (from-to)1877-1886
Number of pages10
JournalDiabetologia
Volume58
Issue number8
Early online date30 May 2015
DOIs
Publication statusPublished - 1 Aug 2015

Keywords

  • GLUT4
  • G-protein photolabel
  • insulin
  • Noc2
  • Rab3
  • Rph3al

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