FIG 9.
PDE and DGC activity assays of DGC14945, PDE14950, and PDE10355. (A to D) HPLC analysis of products in the c-di-GMP PDE enzymatic reaction. Purified PDE10355 and PDE14950 proteins were tested for PDE enzymatic activity using the c-di-GMP molecule as the substrate. (A) pGpG standard. (B) c-di-GMP standard. (C) When incubated with purified PDE10355 for 1 h, c-di-GMP was stable, and no degradation was detected. (D) When incubated with PDE14950 for 10 min, c-di-GMP was hydrolyzed into linear pGpG. (E to H) HPLC analysis of products in the c-di-GMP DGC enzymatic reaction. (E) c-di-GMP standard. (F) GTP standard. (G) GTP was synthesized by WspR into c-di-GMP after incubation for 60 min (positive control). (H) After incubation with purified DGC14945GAF-SGDEF for 7 h, GTP was synthesized into c-di-GMP eventually. DGC14945, PDE14950, and PDE10355 represent W909_14945-, W909_14950-, and W909_10355-encoded proteins, respectively.