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Project: “Analysis of downregulation of MAPK-signaling pathways”Achievements:-first time demonstration of E3 ligase activity of scaffold protein Ste5 in vitro and in vivo-Halo bead assay was first time applied to detect E2-E3 complexes in vitro (weak interaction and high instability of scaffold full-length Ste5 protein hampered application of standard methods)-identification of E2 binding site of Ste5 protein by structural alignment of Ste5 model with published E2-E3 complexes and further supported by experimental data- degradation of the substrate (Ste11) was abolished in presence of the E2–binding deficient Ste5 mutant, same time experimental data confirmed that the general fold of the mutated domain was well-preserved (G-protein binding assays)- for degradation experiments the set of cell lines with no background activity in absence on mating pheromone and no cross-reactivity of overlapping signaling pathways was designed and successfully applied

Project: “Analysis of ubiquitin E3 ligase activity of Ring domain of rhesus Trim5 alpha protein by biochemical and biophysical methods” Achievements: -structural data were gained for three-protein complex (ubiquitin, E3 and E2 proteins) by NMR spectroscopy and revealed the role of dimerization of host restriction factor Trim5alpha for synthesis of ubiquitin K63-linked chains - crystal structure of E2-E3 proteins was solved, explaining the different restriction properties of Trim5alpha mutants in vivo and in vitro -based on results the mechanism of how innate immune response is triggered through the sensing of repetitive viral interface by host restriction factors was revealed -highly cited paper was published with my name as a single first author, experimental part was obtained during less than one year.

Achievements: I had studied two projects in the terms of my PhD studies: 1st project: “Identification of neutralizing epitopes on Dengue virus E protein by NMR spectroscopy”. Achievements: -the epitope regions of envelope protein of Dengue virus in complex with panel of human mAbs was identified by NMR spectroscopy - SPR and mutagenesis experiments conducted to confirm the structural information and antibody-antigen docking simulations were done and filtered in agreement with experimental data 2nd project: “Mapping of neutralizing epitopes on diphtheria toxin by biochemical, immunological and computational methods”. Achievements: -identification of the epitopes, located on diphtheria toxin, done for panel of human monoclonal antibodies -first time demonstration that for toxin neutralization the precise epitope is more important than the binding efficiency of antibody to the antigen - modeling of the antibody 3D-structure (CDR loops regions) based on antibody sequence -antibody-antigen docking simulations to visualize protein complexes were done and filtered based on experimental information (more than 100k complexes were simulated by Rosetta Docking) - PhD thesis defense performed based on the results of the study, the manuscript is on preparation

Achievements: Analysis of lipid packing’s temperature dependence on a membrane model by EPR spectroscopy. I had studied the basics of EPR spectroscopy and applied it to the practical task (lipid packing analysis).

Achievements:Analysis of heme composition of cytochrome c oxidase from Chloroflexus aurantiacus. I was helping to run the set of experiments as a technician working under purification of bacterial membrane proteins (cytochrome c oxidases) for their further analysis (optimization of growth conditions (at microaerofilic environment), solublization, separation of units of the oxidase and optical analysis of their heme composition. As results of this work I had access to all basics methods of biochemistry and gained lively interest to the structural biology and biophysics.