Fig. 3: The pupylation motif on FabD is conformational and not linear. | Nature Communications

Fig. 3: The pupylation motif on FabD is conformational and not linear.

From: Electrostatic interactions guide substrate recognition of the prokaryotic ubiquitin-like protein ligase PafA

Fig. 3

a Cartoon representation of the FabD structure (PDB: 2QC3). Residues around K173 (blue) are colored according to their contribution to the pupylation reactivity of FabD as judged by the effect of the mutation to alanine. Red color indicates a strong contribution manifested as a severe pupylation defect upon mutation. Green colored residues had little effect in the alanine scan and yellow colored residues showed an intermediate effect. b Pupylation time courses recorded by SDS-PAGE and Coomassie-staining for those FabD alanine variants that had the strongest effect on pupylation efficiency. Pupylation time courses for other variants are shown in Supplementary Fig. 4b. Based on these pupylation assays, contribution of individual residues to pupylation efficiency was mapped in a. The band labeled FabD-Pup* was identified by mass spectrometry as FabD pupylated at residues K228 or K291 instead of the native pupylation site K173 (see Supplementary Data 2). Representative gels of three individual experiments are shown. Source Data (uncropped gels) are provided as a Source Data file.

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