Screen of Non-annotated Small Secreted Proteins of Pseudomonas syringae Reveals a Virulence Factor That Inhibits Tomato Immune Proteases
Fig 5
Competitive ABPP screen reveals inhibitor of immune protease C14.
Protein extracts of leaves transiently overexpressing the C14 immune protease were pre-incubated with or without 50 μM E-64 or with 1 μg purified Avr2, EpiC1 or Epic2B (A) or 1 μg purified putative effector protein (B) or for 30 minutes and then incubated for one hour with 1 μM MV201 to label the non-inhibited proteases. Proteins were separated on a 12% protein gel and scanned for fluorescently labeled proteins. The signals were quantified and plotted in a bar graph. The region corresponding to 0.5 to 1.5 times the no-inhibitor-control is shaded grey. This screen was repeated twice with a similar outcome.