FIGURE 2.
Time lapse recording of the SiaDW reaction. A, SiaDW was primed with 1 mm 4-MU-Sia-Gal-Sia (4-MU-DP3) in the presence of 2 mm UDP-Gal and 2 mm CMP-Sia. At the indicated time points, product profiles were recorded by HPLC-FD. Gray peaks, 4-MU and 4-MU-DP1, which were left in the priming 4-MU-DP3 pool. Due to the steep increase in acceptor quality from DP2 to DP3, both peaks remained unchanged over the reaction course. B, enlarged display of the HPLC-FD profile obtained at 40 min and labeling of individual DPs. C, reaction progress curves were generated by quantification of each chromatogram and plotting these values against the reaction time point. Three independent experiments (R1–R3) document the high reproducibility of the reaction.