Figure 8.
GmPDIM oxidizes the active centers of GmPDIL-2. A, The mixed disulfide of the GmPDIM and GmPDIL-2 mutants. GmPDIM(C67A) (6 μm), 3 μm GmPDIL-2(C104/443A), and 1 μm GmERO1a were incubated at 25°C for 60 min and separated by two-dimensional electrophoresis. M1, GmPDIM monomer; L21 GmPDIL-2 monomer; M2, GmPDIM dimer; L2M1 and L2M2, mixed disulfide complex of GmPDIL-2 mutants and GmPDIM(C64A). B, C, D, E, F, Indicated combinations of proteins were incubated and analyzed as described in A. G, H, I , J, K, L, Assay of redox equilibrium constants of GmPDIM, GmPDIL-2, and their mutants. Coomassie Brilliant Blue stainings of the SDS-polyacrylamide gels are shown above the redox graphs. Data are represented as mean ± se of n = 3.