FIGURE 6.

Interaction between the N-terminal cytoplasmic region and the cytosolic C-terminal region, which carries FAD- and NADPH-binding domains. A, domain structures of fragments used in pulldown assays. B, in vitro pulldown assay of MBP-OsRbohB-(590-C) with various N-terminal fragments of thioredoxin-OsRbohB (Trx-OsRbohB). Each N-terminal fragment of OsRbohB was incubated either with maltose binding protein (MBP) or with MBP-OsRbohB-(590-C). The identity of the Trx-OsRbohB-(138–313) band was confirmed by Western blotting using anti-His antibodies. C, effects of Ca²⁺ on the binding. EDTA-treated and E253A mutant of OsRbohB-(1–355) were incubated. No Ca²⁺ effects on the binding were observed. D, in vitro pulldown assays of MBP-OsRbohB containing a cytoplasmic loop with OsRbohB-(1–355).