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Content archived on 2024-05-14

Investigations into the subcellular trafficking of the glut4 glucose transporter in insulin-sensitive tissues

Objective



This project is aimed at understanding how insulin regulates the fusion of intracellular vesicles endowed with the GLUT4-glucose transporter with the plasma membrane. In particular, we aim to identify | proteins involved in the formation of the so called 'fusogenic particle'. To this end, two main approaches will be developed. First, we will carry out in vivo functional assays investigating the effect of introducing, into living cells, inhibitory antibodies against proteins thought to play a key role in membrane fusion. Second, using two recombinant proteins (NSF and a-SNAP), which appear to be essential components of the intracellular membrane fusion apparatus, an affinity purification procedure (based on the natural binding of these proteins to their targets) will be developed to isolate proteins from immunopurified GLUT4-containing vesicles which are involved in the formation of the fusogenic machinery. Training content (objective, benefit and expected impact) The applicant will be trained in a variety of methods including generation of polyclonal and monoclonal antibodies, protein over-expression in Echerichia coli and purification of recombinant proteins, cell culture and protein purification. Other techniques include l electroporation of cells and subcellular fractionation. ; It is our belief that we, as a group, are suitably poised to address the questions raised in the attached application and that information generated from the proposed studies will contribute significantly to our conceptual understanding of how glucose transport is regulated during physiological and pathological I circumstance. Links with industry / industrial relevance (22)

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Programme(s)

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Topic(s)

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Call for proposal

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Funding Scheme

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Coordinator

Aristotle University of Thessaloniki
EU contribution
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Address
Analytical Chemistry Lab
54006 Thessaloniki
Greece

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Total cost

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