Protein kinase-ζ interacts with munc18c: role in GLUT4 trafficking
Hodgkinson, C.P., Mander, A. and Sale, G.J. (2005) Protein kinase-ζ interacts with munc18c: role in GLUT4 trafficking. Diabetologia, 48, (8), 1627-1636. (doi:10.1007/s00125-005-1819-y).
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Aims/hypothesis: Insulin-stimulated glucose transport requires a signalling cascade through kinases protein kinase (PK) Cζ/λ and PKB that leads to movement of GLUT4 vesicles to the plasma membrane. The aim of this study was to identify missing links between the upstream insulin-regulated kinases and the GLUT4 vesicle trafficking system.
Materials and methods: A yeast twohybrid screen was conducted, using as bait full-length mouse munc18c, a protein known to be part of the GLUT4 vesicle trafficking machinery.
Results: The yeast twohybrid screen identified PKCζ as a novel interactor with munc18c. Glutathione S transferase (GST) pull-downs with GST-tagged munc18c constructs confirmed the interaction, mapped a key region of munc18c that binds PKCζ to residues 295–338 and showed that the N-terminal region of PKCζ was required for the interaction. Endogenous munc18c was shown to associate with endogenous PKCζ in vivo in various cell types. Importantly, insulin stimulation increased the association by approximately three-fold. Moreover, disruption of PKCζ binding to munc18c by deletion of residues 295–338 of munc18c or deletion of the N-terminal region of PKCζ markedly inhibited the ability of insulin to stimulate glucose uptake or GLUT4 translocation.
Conclusions/interpretation: We have identified a physiological interaction between munc18c and PKCζ that is insulin-regulated. This establishes a link between a kinase (PKCζ) involved in the insulin signalling cascade and a known component of the GLUT4 vesicle trafficking pathway (munc18c). The results indicate that PKCζ regulates munc18c and suggest a model whereby insulin triggers the docking of PKCζ to munc18c, resulting in enhanced GLUT4 translocation to the plasma membrane.
|Keywords:||glucose transport, GLUT4, insulin, munc18c, protein kinase C|
|Subjects:||R Medicine > RB Pathology
Q Science > QH Natural history > QH301 Biology
|Divisions:||University Structure - Pre August 2011 > School of Biological Sciences
|Date Deposited:||03 Apr 2006|
|Last Modified:||28 Jun 2012 10:05|
|Contributors:||Hodgkinson, C.P. (Author)
Mander, A. (Author)
Sale, G.J. (Author)
|Contact Email Address:||G.J.Sale@soton.ac.uk|
|RDF:||RDF+N-Triples, RDF+N3, RDF+XML, Browse.|
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