Fig. 4: Molecular mechanism of FBP-PGAM1 interaction through phosphate transfer.

a LC-MS/MS detection of F6P and F1P generation at different time slots after FBP treatment of purified PGAM1. b Schematic illustration of the phosphate transfer from FBP to PGAM1, generating either F6P upon C1-O-phosphate donation or F1P upon C6-O-phosphate donation. c Molecular docking reveals the hydrogen bonding network between FBP and PGAM1 when it donates either its C1-O-phosphate or its C6-O-phosphate. d Immunoblotting analysis and dissociation constants of PGAM1 mutants PGAM1-R10A, PGAM1-R62A, PGAM1-E89A, PGAM1-Y92A, PGAM1-H186A PGAM1-N188A and PGAM1-3A (R10A, E89A and H186A) with key residues involved in the FBP-PGAM1 binding mutated (n = 3 independent experiments with similar results). e Steady state of PGAM1–FBP complex during the process of phosphate transfer from FBP to PGAM1 His11 after 750 ns MD simulation and the probabilities of hydrogen bonding interactions between FBP and different PGAM1 active site residues during the process of either C1-O-phosphate transfer (up) or C6-O-phosphate transfer (down). Orange represents attractive charges, blue represents conventional hydrogen bonds. Size represents the probability of hydrogen bonding interaction between FBP functional group and amino acid residue within the PGAM1 active site. All measurements are presented as mean ± SD from three biological replicates. Source data are provided as a Source Data file.