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The reaction of TycC TE with IT1-SNAC followed by HPLC. Reactions initially contained 15 μM IT1-SNAC, 200 nM TycC TE, 25 mM Mops, pH 7.0 at 24°C and were quenched at time 0 min or 10 min. The labeled hydrolyzed and cyclic products, IT1-COOH and cyclo-IT1, respectively, were identified by MALDI-TOF MS. (Inset) Shown are peptide fragment ions generated by MS-MS that confirm the presence of the underlined d-Phe-1 to Leu-10 linkage formed by cyclization.
