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Information × Registration Number 0222U000386, 0120U104648 , R & D reports Title The nonproteinogenic leucine analogues in leucine-induced mTORC1 signaling in human cells popup.stage_title Head Tukalo Mykhailo A., Доктор біологічних наук Registration Date 07-01-2022 Organization Institute of Molecular Biology and Genetics of NAS of Ukraine popup.description2 The effect of leucine analogues on the phosphorylation of S6K1 and ULK1 kinases in HEK293T cells was determined. Non-proteinogenic leucine analogues were shown to stimulate phosphorylation of ULK1, while S6K1 activity remained unaffected. Codon-optimized RagB and RagD GTPases were cloned into bacterial vector for coexpression into E.coli cells. Site-directed mutagenesis of RagB and RagD was performed. RagA/RagC, FLCN-FNIP2 and Sestrin2 were purified. The conditions for Sestrin-GATOR2 purification were elaborated. Complex assembling for human cytosolic leucyl-tRNA synthetase (HscLeuRS), Rags and FLCN-FNIP2 was studied by in vitro assay and molecular dynamics simulation methods. The formation of complexes was shown to depend on nucleotide-bound state of Rags but not to depend on leucine or leucyl-adenylate-bound state of HscLeuRS.   Product Description popup.authors Areshkov Pavlo O. Zhuk Oleksandr V. Kovalenko Oksana P. Kochutenko Alona V. Rayevsky Olexey V. Skydanovych Oleksandra I. Tukalo Mykhailo А. Shablii Volodymyr A. popup.nrat_date 2022-03-09 Close