Fig 5. T6SS-4 translocates a Zn2+-binding protein.
A. The binding of zinc ions by YezP. Zn2+-free YezP (upper) or YezPH76A (lower) was used to evaluate zinc-binding activity by isothermal titration calorimetry (ITC). Data were analyzed with the NanoAnalyze software (TA Instruments). B. YezP is a secretion substrate of T6SS-4. Proteins in culture supernatant of relevant Yptb strains expressing YezP-VSVG were probed for VSVG (upper) or Hcp4-VSVG (lower) by immunoblotting. For the pellet fraction, the isocitrate dehydrogenase (ICDH) was detected as loading controls. Similar results were obtained in three independent experiments, and data shown are from one representative experiment done in triplicate.