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To generate the respective peptide library initiated with ClAc-L-Tyr (L-library) or ClAc-D-Tyr (D-library), 1.4 M mRNApuromycin library was translated in a methionine-deficient FIT system 14 at a scale of 150 l total volume in the presence of 50 M ClAc-L-Tyr-tRNA fMet CAU or ClAc-D-Tyr-tRNA fMet CAU for 30 min at 37 C
In each case, tandem MS analysis showed neutral losses of fragments of 75 Da, 129 Da, and 307 Da, along with an m/z 308 ion, characteristic of glutathione conjugates (Supplementary Figure S1)