Fig. 4: Characterization of LctF and LctA_AT1. | Communications Chemistry

Fig. 4: Characterization of LctF and LctA_AT1.

From: Biosynthesis of lactacystin as a proteasome inhibitor

Fig. 4

a LC-ESI-MS analysis (negative ion mode, monitored at m/z 794) of (i) formyl-CoA standard and reaction mixtures containing (ii) LctF and 10N-fTHF, (iii) 10N-fTHF, and (iv) LctF. b LC-MS analysis of reaction mixtures of LctA_AT1 and formyl-CoA; (i) without LctA_AT1, (ii) with LctA_AT1 (Fig. S8). A mass of 12794.3 corresponding to formyl-ACP1 (calculated mass, 12793.5) was detected in (ii). c LC-MS analysis of reaction mixtures of a triple mutant of LctA_AT1 (M-LctA_AT1) with (i) acetyl-CoA and (ii) formyl-CoA (Fig. S9). A mass of 12807.9 corresponding to acetyl-ACP1 (calculated mass, 12807.5) was detected in (i). d Relative activities of LctA_AT1 and M-LctA_AT1 toward formyl- (For-), acetyl- (Ac-), and malonyl- (Mal-) CoA. The activities were quantified with Ellman’s reagent (DTNB) reacting with free thiol released from the substrates. Error bars indicate standard deviation (SD) (n = 3), and data are presented as mean values ± SD. Black dots denote individual data points.

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