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The project is aimed at the study of molecular and functional properties of unique proteins affecting haemostasis, angiogenesis, apoptosis. The proteins will be separated from viper venoms (Levantine viper, Common viper) combining protein purification methods such as gel filtration, ion-exchange chromatography, HPLC, affinity chromatograpy, electrophoresis and isoelectric focusing. The protein interactions with cells will be detected using platelets, human umbilical vein endothelial cells (HUVEC), K-562 and some other tumor cells. The primary structure of the proteins separated from Levantine viper (disintegrin/disintegrin-like proteins, apoptosis inducing metalloproteinases) will be deduced from the DNA encoding the protein using our Levantine viper venom gland cDNA library.The analysis of obtained protein structures and literature provided data allows to elucidate structure-function relationships and also the peculiarities of the binding of these proteins to different cell surface receptors. We plan to obtain disintegrin/disintegrin-like (usually nonglycosylated) recombinant proteins using bacterial expression systems. The molecular and functional properties of recombinant proteins will be compared with native ones.The results of the project will be essential for protein-protein interaction fundamental studies. Inhibitors of angiogenesis and platelet aggregation may be potential antitumor and antithrombotic drugs.The structural studies are useful for designing more effective potential drugs. |