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For binding assays, the purified proteins were incubated with glutathione high-capacity magnetic agarose beads in binding buffer (50 mM Tris-HCl, 150 mM KCl, 5 % glycerol, 1 mM DTT, 1 mM EDTA, pH 8.0, 0.01 % Nonidet P-40, 1 mM PMSF) at 4 C for 3 h

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Published 2017 Dec 22 Tahara Y, Hirota M, Shima K, et al

designed a prodrug form (JHU083) of the glutamine antagonist DON, administered in an inert state but then preferentially activated by enzymes enriched in the tumor microenvironment [72]