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. 2018 Sep 27;9:2347. doi: 10.3389/fmicb.2018.02347

FIGURE 2.

FIGURE 2

Phosphodiesterase activity of rGdpP109-657-ProS2 and rDhhP-ProS2 by liquid chromatography (LC) analysis. Reaction mixtures (50 μl) contained 50 mM potassium phosphate buffer (pH 5.0 for GdpP, pH 7.0 for rDhhP-ProS2), 1 mM MnCl2, 100 μM substrate (c-di-AMP or pApA), and 1.2 μg ml-1 rGdpP109-657-ProS2 or 0.36 μg ml-1 rDhhP-ProS2. Reactions were incubated for 15 min and terminated by addition of TE-saturated phenol and chloroform, and an aliquot (1 μl) of the aqueous phase was injected onto the LC column. Each sample was separated by LC and monitored at 254 nm. Purified AMP, c-di-AMP, and pApA were used as standards.