Fig. 4
From: Functional role of PGAM5 multimeric assemblies and their polymerization into filaments
PGAM5 multimerization is required for catalysis. a Overview of interactions contributing to the stability of the dodecamer assembly in the structure of ∆90 PGAM5 H105A/MM. b SEC elution profiles of wild type (WT) and interface-disrupting mutants (R65A, H105A, Y198E, F244E, and A4) of ∆48 PGAM5 in 150 mM NaCl-containing buffer, highlighting the effect of the MM (R65A, A4), and dimer interface (F244E, Y198E) mutants on the oligomer/dimer equilibria observed for wild-type ∆48 PGAM5. c EM micrographs of negatively stained samples of interface mutants of PGAM5 corresponding to the oligomer peaks for WT and H105A, and dimer/monomer peaks for all the other mutants as shown in b. The insets in top right corners illustrate the position of the respective mutations within the PGAM5 dodecamer cartoon. Scale bars correspond to 50 nm. d Phosphatase activity of the WT and mutant variants of ∆48 PGAM5 measured for 20 nM enzyme and varying concentrations of a phosphorylated ASK1 substrate peptide. Data are represented as mean ± S.E.M. The kinetic parameters Km, kcat, and kcat/Km were determined using GraphPad Prism and are summarized in Table 2
