Gabin de la Bourdonnaye
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Background: Training and Transferable Skills:
Research Projects: RATIONAL ENGINEERING AND BIOPHYSICAL CHARACTERIZATION OF FIBROBLAST GROW FACTORS The fibroblast growth factors (FGFs) are extracellular proteins that regulate many aspects of embryonic development and adult homeostasis. FGFs are involved in angiogenesis, wound healing, and embryonic development regulating growth of specific tissues. These proteins have great potential for use in medicine, cosmetics and stem cell research. However, the application potential of FGFs is limited due to their natural instability. Main goal of this project will be rational and semi-rational engineering of selected FGF proteins to enhance their thermodynamic and kinetic stability. Stabilized proteins will be subsequently characterized to determine their binding interactions with heparan sulfate by using of various biophysical methods. The FGF biological functions are controlled by their interactions with heparan sulfate. Transport of FGFs between cells is determined by heparan sulfate (co-receptor) binding which is required for the assembly of high affinity signaling complexes with their cognate FGF receptor. Moreover, the interactions of FGFs with heparan sulfate in extracellular matrices control their transport from a source to a target cell. Various methods of molecular biology including site directed mutagenesis, site saturation mutagenesis and directed evolution will be used to construct stable variants of selected FGFs. Diversification of the specificity of FGF-heparan sulfate interactions will be identified by a range of complementary techniques, such as isothermal titration calorimetry, circular dichroism spectroscopy and differential scanning fluorimetry. Biological activity of evolved molecules will be evaluated by the CyQUANT® Cell Proliferation Assays with cell lines of mouse fibroblast (e.g., NIH/3T3). |
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