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Information × Registration Number 0220U100488, 0115U004200 , R & D reports Title Modeling of molecular processes underlying Parkinson's disease in yeast cells and analyzing the impact of exogenous and endogenous factors on the processes of aggregation and degradation of human alpha synuclein popup.stage_title Head Stasyk Oleh V., Кандидат біологічних наук Registration Date 20-01-2020 Organization Institute of Cell Biology popup.description2 The object of the study is the role of arginine deficiency in the degradation of heterologous neuronal alpha-synuclein protein in cells of the H. polymorpha model strain. The purpose of the research phase is to investigate the molecular mechanisms of degradation of abnormal forms of human alpha-synuclein in the model microorganism H. polymorpha due to arginine amino acid deficiency. Research Methods - Methods for fluorescence analysis (detection of chimeric forms of α-synuclein conjugated to green fluorescent protein (GFP)); microbiological methods (growth kinetics analysis of model strains); molecular biological methods (Western blot detection of α-synuclein protein using murine anti-α-synuclein antibodies); apparatus (polyacrylamide gel protein electrophoresis cameras, device for semi-wet protein transfer to nitrocellulose or PVDF membrane; fluorescence microscope AxioScope.A1, shaker for cultivation of Double Deck Incubator yeast culture. As a result of the work, strains of the methylotrophic yeast Hansenula (Ogataea) polymorpha were first designed to produce human alpha-synuclein labelled with a green fluorescent protein (EGFP). To simulate the effect of arginine deficiency on the processes of autophagic degradation of alpha-synuclein, which can be generated in vivo using arginine-degrading enzymes, the gene of argininosuccinate synthetase was deleted from the model strain. Arginine deficiency was found to induce autophagy in the cells of the auxotrophic model strain and reduce its intracellular content but did not cause complete degradation of the heterologous protein. Thus, the deficiency of individual amino acids can be used to treat Parkinson's disease and the study of its effects should be continued in alternative laboratory models. Product Description popup.authors Vovk Olena I. Maievska Oksana M. Stasyk Oleh V. popup.nrat_date 2020-04-02 Close