Fig. 5: Reduction of disulfide bond in CadC by DsbC. | Communications Biology

Fig. 5: Reduction of disulfide bond in CadC by DsbC.

From: Molecular mechanism of proteolytic cleavage-dependent activation of CadC-mediated response to acid in E. coli

Fig. 5

qRT-PCR analysis of cadA (a) and cadB (b) gene expression in MG1655ΔdsbC, ΔdsbG, and ΔlysP strains following acid and lysine stimulation. Data are mean ± SD of n = 3 biologically independent replicates. Different letters denote significant differences among various samples at the same detection time, according to Duncan’s test (p < 0.05, n = 3). c LDC assay was used to assess lysine decarboxylation capacity in MG1655, Δ dsbC, Δ dsbG, and Δ lysP strains. d qRT-PCR analysis of cadA and cadB gene expression in MG1655ΔdsbC, MG1655ΔdsbC-pcadC C208S, MG1655ΔdsbC-pcadC C272S, and MG1655ΔdsbC-pcadC C172S. Data are mean ± SD of n = 3 biologically independent replicates. e, f Western blot assay examining the protein levels of CadC in MG1655ΔdsbG, MG1655ΔdsbC, and MG1655ΔlysP strains. DnaK served as a control for immunoblotting analysis of total protein. g, qRT-PCR was used to detect the expression of cadBA gene in mutant strains MG1655ΔlysP: cadC-pcadC C208S and MG1655ΔlysP: cadC-pcadC C272S. Data are mean ± SD of n = 3 biologically independent replicates. h Western blot assay examining the protein levels of CadC in MG1655ΔlysP: cadC-pcadC C208S, MG1655ΔlysP: cadC-pcadC C272S strain. DnaK served as a control for immunoblotting analysis of total protein. i A spot assay on low pH agar is performed to evaluate the survival/growth of MG1655, MG1655ΔcadC-pcadC C208S, MG1655ΔcadC-pcadC C272S, MG1655ΔcadC, MG1655ΔcadC-pcadC R184Q, MG1655ΔcadC-pcadC; MG1655ΔdegP: degQ, MG1655ΔdegP, MG1655ΔdegQ, MG1655ΔlysP, MG1655ΔdsbC; MG1655ΔlysP: cadC-pcadC C208S, MG1655ΔlysP: cadC-pcadC C272S strains.

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