Gorbenko O. Identification and characterization of PTEN phosphatase binding proteins

The object - molecular mechanisms of cell signal system function. The aim of the work was to reveal a novel functional links between PTEN phosphatase and proteins that involved in different signal pathways by yeast two-hybrid system and to confirm the PTEN-novel protein interaction found in yeast using methods of molecular biology in vitro and the systems of high eukaryotes. Applied methods - cDNA fragment cloning and site-directed mutagenesis, yeast two-hybrid screen, yeast mating assay, surface plasmon resonance BIAcore, GST-pull down assay and gel-filtration. Results and novelty - new bait-constructs expressing C-terminal and full mutant PTEN have been created. The screening of cDNA Colon Cancer, 18-day Mouse Embryo and HeLa cell libraries has been carried out. Eight new PTEN-binding proteins have been revealed and the specificity of the interactions was confirmed in yeast cells. The monoclonal anti-FABP4 antibodies have been generated. The possibility of PTEN-FABP4 complex formation in vitro and inadipocytes has been shown. The hypothesis of PTEN possible function in adipocytes has been proposed.