The vascular endothelial growth factor receptor 2 (VEGFR2) plays a key role in angiogenesis and vascular physiology in higher eukaryotes such as vertebrates. Although intracellular signalling pathways linked to VEGFR2 activation have been well characterised, there is a lack of knowledge on membrane and cytosolic effectors that interact with VEGFR2. To address this problem, I have used a novel membrane protein yeast 2-hybrid (Y2H) screen to identify novel factors that interact with membrane VEGFR2. By screening a human primary endothelial cDNA library, six potential VEGFR2-binding proteins were isolated. One candidate was the SlOO calcium-binding protein A6 (SlOOA6). SlOOA6 is an abundant protein present in both the nucleus and cytoplasm of endothelial cells. The interaction between VEGFR2 and SlOOA6 was found to be calcium-dependent. Calcium dependent-translocation of S lOOA6 from the nucleus caused eo-distribution with VEGFR2 in punctate vesicles. Depletion of SI OOA6 protein levels using RNA interference perturbed VEGF-A-stimulated VEGFR2 activation and phosphorylation, cell proliferation, cell migration and angiogenesis. Taken together, the work in this thesis suggests that S lOOA6 is a novel binding partner for VEGFR2 and regulates intracellular signalling and angiogenesis. Thus SlOOA6 could be a new target in the treatment of vascular disease.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:555858 |
| Date | January 2011 |
| Creators | Bao, Leyuan |
| Publisher | University of Leeds |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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