title: Insulin regulates Rab3-Noc2 complex dissociation to promote GLUT4 translocation in rat adipocytes.
abstract: OBJECTIVE
  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
  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.
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, BA2 7AY, UK.
authors.forename: Francoise
authors.initials: F
authors.lastname: Koumanov
authors.forename: Vinit J
authors.initials: VJ
authors.lastname: Pereira
authors.forename: Judith D
authors.initials: JD
authors.lastname: Richardson
authors.forename: Samantha L
authors.initials: SL
authors.lastname: Sargent
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Fazakerley
authors.forename: Geoffrey D
authors.initials: GD
authors.lastname: Holman
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism; D000818:Animals; D051275:Glucose Transporter Type 4 / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology*; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D051379:Mice; D021381:Protein Transport / Q000187:drug effects; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects; D020692:rab3 GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D051275:Glucose Transporter Type 4; D007328:Insulin; D047908:Intracellular Signaling Peptides and Proteins; D011506:Proteins; C109307:Rph3al protein, rat; C495694:Slc2a4 protein, rat; D020692:rab3 GTP-Binding Proteins