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In order to extract second-order rate constants, the model thioester and the Cys peptides were reacted at a respective concentration of 0.1 and 0.15 mM (in phosphate buffer 0.1 M, 6 M Gn.HCl, 200 mM MPAA, 100 mM TCEP, pH 7.2, 37 C)

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