Fig. 6: Characterization of ISA1-ISA2 heterocomplex from maize, potato and Arabidopsis.

Gel filtration analysis of ISA1, ISA2, and ISA1-ISA2 heterocomplex of maize (a), Arabidopsis (b) and potato (c). Specific tags and boundaries are shown on top. The signal peptide was removed for protein expression. d–f Protein mass weight examined by analytical ultracentrifugation characterization. The values of molecular weight are shown. g, h Protein oligomerization examined by native PAGE analysis. The left panel indicates samples visualized by Coomassie blue staining. The right panel indicates samples visualized by silver solution staining. A total of 2.5 μL 50 nM protein was used for each sample. i Enzymatic activity of ISA1, ISA2, and ISA1-ISA2 complex of maize, Arabidopsis and potato. 4 nM ISA1, 4 nM ISA2, and 1 nM ISA1-ISA2 complex were used. At least three biological replicates were performed. Data are presented as mean values +/− SEM (n = 9; *p < 0.05; **p < 0.01; ***p < 0.001; unpaired t-test, error bars = mean ± SEM).