Extended Data Fig. 4: PPIaseGS077 is the target of BfUbb.
a, b, Size-exclusion chromatography analyses of BfUbb in the presence of PPIaseGS077 (a) or PPIaseGS084 (b). Purified BfUbb was incubated with purified PPIaseGS077 or PPIaseGS084 and subsequently analysed using size-exclusion chromatography in a Superdex 75 increase column. Elution profiles along with SDS-PAGE analyses of the elution fractions are shown. This experiment was conducted at least three times with consistent results. c, d, f, The binding affinities of BfUbb (c and d) and BfUbbC70V (f) to PPIaseGS077 (c and f) or PPIaseGS084 (d) were measured by ITC. The N values were fixed to 1 during the data fitting for d and f. The Kd and ΔH values data are representative of 2-3 independent replicates. e, Confocal microscopy views of B. fragilis GS077S treated with BfUbb, BfUbbC70V, and BfUbbC76G after 5 hours, respectively. Cellular membranes are stained with lipophilic dye FM 1-43 (green); scale bar, 5 μm; the images represent at least 20 images from three independent experiments.
