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Protein kinase-zeta interacts with munc18c: role in GLUT4 trafficking

Protein kinase-zeta interacts with munc18c: role in GLUT4 trafficking
Protein kinase-zeta interacts with munc18c: role in GLUT4 trafficking
AIMS/HYPOTHESIS: Insulin-stimulated glucose transport requires a signalling cascade through kinases protein kinase (PK) Czeta/lambda 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 two-hybrid 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 two-hybrid screen identified PKCzeta 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 PKCzeta to residues 295-338 and showed that the N-terminal region of PKCzeta was required for the interaction. Endogenous munc18c was shown to associate with endogenous PKCzeta in vivo in various cell types. Importantly, insulin stimulation increased the association by approximately three-fold. Moreover, disruption of PKCzeta binding to munc18c by deletion of residues 295-338 of munc18c or deletion of the N-terminal region of PKCzeta markedly inhibited the ability of insulin to stimulate glucose uptake or GLUT4 translocation.
CONCLUSIONS/INTERPRETATION: We have identified a physiological interaction between munc18c and PKCzeta that is insulin-regulated. This establishes a link between a kinase (PKCzeta) involved in the insulin signalling cascade and a known component of the GLUT4 vesicle trafficking pathway (munc18c). The results indicate that PKCzeta regulates munc18c and suggest a model whereby insulin triggers the docking of PKCzeta to munc18c, resulting in enhanced GLUT4 translocation to the plasma membrane.
0012-186X
1627-1636
Hodgkinson, C.P.
d7d2d1d8-5e1f-4074-a60b-14c38fdc6ad0
Mander, A.
2a92e8dd-4bb4-4f3b-b8ca-d62f5c4724bd
Sale, G.J.
9da2ccd1-1a75-42e8-8c55-5bdfef6ccd10
Hodgkinson, C.P.
d7d2d1d8-5e1f-4074-a60b-14c38fdc6ad0
Mander, A.
2a92e8dd-4bb4-4f3b-b8ca-d62f5c4724bd
Sale, G.J.
9da2ccd1-1a75-42e8-8c55-5bdfef6ccd10

Hodgkinson, C.P., Mander, A. and Sale, G.J. (2005) Protein kinase-zeta interacts with munc18c: role in GLUT4 trafficking. Diabetologia, 48 (8), 1627-1636.

Record type: Article

Abstract

AIMS/HYPOTHESIS: Insulin-stimulated glucose transport requires a signalling cascade through kinases protein kinase (PK) Czeta/lambda 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 two-hybrid 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 two-hybrid screen identified PKCzeta 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 PKCzeta to residues 295-338 and showed that the N-terminal region of PKCzeta was required for the interaction. Endogenous munc18c was shown to associate with endogenous PKCzeta in vivo in various cell types. Importantly, insulin stimulation increased the association by approximately three-fold. Moreover, disruption of PKCzeta binding to munc18c by deletion of residues 295-338 of munc18c or deletion of the N-terminal region of PKCzeta markedly inhibited the ability of insulin to stimulate glucose uptake or GLUT4 translocation.
CONCLUSIONS/INTERPRETATION: We have identified a physiological interaction between munc18c and PKCzeta that is insulin-regulated. This establishes a link between a kinase (PKCzeta) involved in the insulin signalling cascade and a known component of the GLUT4 vesicle trafficking pathway (munc18c). The results indicate that PKCzeta regulates munc18c and suggest a model whereby insulin triggers the docking of PKCzeta to munc18c, resulting in enhanced GLUT4 translocation to the plasma membrane.

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Published date: 1 August 2005

Identifiers

Local EPrints ID: 56622
URI: https://eprints.soton.ac.uk/id/eprint/56622
ISSN: 0012-186X
PURE UUID: a9b3de4b-c1b9-452b-abb1-7bbfb0e5d1f7

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Date deposited: 08 Aug 2008
Last modified: 17 Jul 2017 14:30

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