Figure 4.

WA selective covalently binds to the Cys295 residue of PHGDH. (A) The recombinant PHGDH incubated without (up) or with (down) WA, then the peptide fragments were identified by MS/MS. ca, carbamidomethylation, 57.02 Da. (B) The recombinant PHGDH WT and PHGDH mutant proteins were incubated with WP, and followed by protein affinity pull-down assay. The PHGDH bound to the WP were detected by immunoblotting. (C) Dose–response inhibition curve of PHGDH enzyme activity by WA with the recombinant Cys295A PHGDH protein. Data are presented as the mean ± SD, n = 6. (D) BLI analysis of interaction between WA and recombinant Cys295A PHGDH protein.