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Information × Registration Number 0219U102043, 0119U102065 , R & D reports Title Salicylic acid binding proteins in plants: study by biochemical methods and by molecular dynamics popup.stage_title Head Kravets Volodymyr S., Доктор біологічних наук Registration Date 16-12-2019 Organization V.P. Kukhar Institute of Bioorganic Chemistry and Petrochemistry of the National Academy of Sciences of Ukraine popup.description2 The recombinant GAPDH A1 protein was obtained in pure form (> 90%) and further purified by dialysis with subsequent concentration using Amicon Ultra Centrifugal Filter Unit columns. GAPDH A1 R81L mutant protein was prepared in a similar manner. Based on data from computer simulations and molecular simulations using GROMACS, this protein may have a reduced ability to bind salicylic acid. The actual level of binding to salicylic acid of these proteins was tested by surface plasmon resonance technique. Immobilized 3-aminoethyl salicylic acid was used as the ligand. The native GAPDH A1 protein has been shown to bind immobilized 3-aminoethyl salicylic acid with high affinity - the dissociation constant of this interaction was Kd = 28.3 nM. Additionally, a study was conducted on competitive binding of immobilized 3-aminoethyl salicylic acid in the presence of excess free salicylic acid. It has been shown that the presence of 2mM of free salicylic acid leads to a virtually complete loss of the interaction of GAPDH A1 with the immobilized ligand. These results indicate that two compounds compete for the same binding site on the protein surface. In turn, the level of competition for binding to 4-hydroxybenzoic acid (a biologically inactive salicylic acid isomer) was significantly lower. These data indicate the biological significance of the GAPDH A1-salicylic acid interactions. Product Description popup.authors Pokotylo Igor V. popup.nrat_date 2020-04-02 Close