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. 2019 Mar 14;17(8):1670–1678. doi: 10.1111/pbi.13092

Figure 5.

Figure 5

Sl CYS8 inhibits different isoforms of granulin‐containing proteases. (a) Leaf extracts from agroinfiltrated plants overexpressing C14, the tomato RD21‐like protease, with or without Sl CYS8 were labelled with MV201 to monitor PLCP activity. In‐gel fluorescence confirmed inhibition of C14 upon Sl CYS8 co‐expression, while no effect was observed when co‐expressed with the inactive Q47 P S l CYS8 mutant. As a control, plant extracts were mixed and pre‐incubated with E‐64 inhibitor before incubation with or without (Ctrl) the probe. (b) Immunoblot shows equal amount of the intermediate protease (iC14) and the mature (mC14) form of C14 protease. (c) Coomassie gel shows that protein degradation occurred upon C14 expression and is prevented by co‐expressing Sl CYS8. The large subunit of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (RbcL) is stabilized in Sl CYS8‐expressing leaves.