Drobot L. 3. Features of signalling network organization and functioning in normal and tumour cells

3. The objeсt: signalling networks of normal and tumour cells. The aim: to elucidate features of signalling processes dynamics in normal and tumour cells, the role of adaptor/scaffold proteins stechiometry providing for specificity and intensity of intracellular signalling. Methods: construction of expression vectors, expression of recombinant proteins in bacterial, baculoviral and mammalian systems, production of polyclonal and monoclonal antibodies, Western and Nothern blot analysis, immunofluorescense microscopy, immunohistochemistry, purification of proteins using ion exchange and affinity chromatography, determination of enzymatic activity of a set of signalling proteins. Theoretical and practical results: a range of experimental studies have been carried out by results of which the hypothesis about system character of signalling processes oscillations in normal and tumour cells as well as their role in coordination of coupled signalling pathways was suggested. Conception regarding the critical role of adaptor/scaffold proteins stechiometry providing for specificity and intensity of intracellular signalling was suggested and experimentally proved. The regulatory subunit of phosphatidylinositol 3-kinase and endonuclease(es) were shown to be a novel binding partners of adaptor protein Ruk. Decrease of expression of adaptor protein Ruk/CIN85 was established in human kidney light cell carcinomas. The obtained data validate the possibilities of signalling targeted therapy under human pathological states.