FIG. 2.

Analysis of the formation of products from the reaction of PGH with PABA-GLU. Reactions containing 50 mM phosphate buffer, pH 8.5, 2 mM MnCl₂, and 1 mM PABA-GLU were initiated by the addition of purified PGH. At various time points, samples (500 μl) were quenched and analyzed for PABA as described in the text. A parallel sample (600 μl) was inactivated by placement in a boiling water bath for 1 min; enzyme was removed using a Microcon YM-10 filter device with a 10,000-molecular-weight cutoff (Millipore, Billerica, MA), and 400 μl of this sample was subjected to dabsylation and analysis by HPLC as described in the text. Samples were done in triplicate, repeated three times, and analyzed using GraphPad Prism 5. The data shown are averages plus standard errors (SE).