Journal: International Journal of Molecular Sciences

Article Title: Novel Drug-like HsrA Inhibitors Exhibit Potent Narrow-Spectrum Antimicrobial Activities against Helicobacter pylori

doi: 10.3390/ijms251810175

Figure Lengend Snippet: Antimicrobial activities of relevant HsrA inhibitors against C. jejuni and several representative members of normal human microbiota.

Article Snippet: Likewise, a 420-bp sequence of the C. jejuni sodB promoter (ATCC 33560) was used as the target DNA of CosR [ ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Novel Drug-like HsrA Inhibitors Exhibit Potent Narrow-Spectrum Antimicrobial Activities against Helicobacter pylori

doi: 10.3390/ijms251810175

Figure Lengend Snippet: HsrA ligand V noticeably inhibited the in vitro DNA binding activity of the C. jejuni response regulator CosR, an ortholog protein of HsrA. Recombinant CosR was mixed with its target promoter P sodB in the presence of 2, 1, 0.5, and 0.1 mM of ligand V. DMSO instead of the inhibitor was included as the vehicle control, while an internal sequence of the Anabaena sp. pkn22 gene was used as a non-specific competitor in all assays. The addition of protein in the reaction mixtures is indicated by +. Protein–DNA interactions were analyzed by 6% non-denaturing electrophoresis.

Article Snippet: Likewise, a 420-bp sequence of the C. jejuni sodB promoter (ATCC 33560) was used as the target DNA of CosR [ ].

Techniques: In Vitro, Binding Assay, Activity Assay, Recombinant, Control, Sequencing, Electrophoresis

Journal: Microbial Biotechnology

Article Title: Additive effects of metal excess and superoxide, a highly toxic mixture in bacteria

doi: 10.1111/1751-7915.13589

Figure Lengend Snippet: R. gelatinosus SodB expression and activity. (A) Western blot analysis of wild‐type ( WT ) and Δ sodB mutant cells. Cells were grown by photosynthesis and total protein extracts from the same amount of cells (0.1 OD 680nm ) were analysed by Western blot. (B) Involvement of SodB in oxidative stress under photosynthetic (PS) or respiratory (RES) condition. Representative growth inhibition (left) and mean ± stdev from 3 independent experiments of the diameter of inhibition zone (right) are presented. (C) SOD in‐gel activity assay of indicated strains. Expression and activity profile of SodB in the WT , Δ cadA strain after various CdCl 2 treatment of cells grown under PS (D) and RES (E) conditions.

Article Snippet: In the cyanobacterium Synechocystis sp. PCC 6803, high copper treatment also induced the expression of the iron superoxide dismutase sodB (Giner‐Lamia et al. , ).

Techniques: Expressing, Activity Assay, Western Blot, Mutagenesis, Inhibition

Journal: Microbial Biotechnology

Article Title: Additive effects of metal excess and superoxide, a highly toxic mixture in bacteria

doi: 10.1111/1751-7915.13589

Figure Lengend Snippet: Involvement of SodB in Cd 2+ stress in R. gelatinosus . Dose–response curves of the wild‐type ( WT ), ΔsodB , ΔcadA and ΔcadAsodB strains. Cells were grown for 24 h in the presence of different concentrations of CdCl 2 either aerobically (A) or by photosynthesis (B). Results represent the mean of 3 independent experiments. Menadione disc diffusion assay of cells in RES condition with or without 25 µM CdCl 2 (C). Results represent mean ± stdev from 3 independent experiments.

Article Snippet: In the cyanobacterium Synechocystis sp. PCC 6803, high copper treatment also induced the expression of the iron superoxide dismutase sodB (Giner‐Lamia et al. , ).

Techniques: Diffusion-based Assay

Journal: Microbial Biotechnology

Article Title: Additive effects of metal excess and superoxide, a highly toxic mixture in bacteria

doi: 10.1111/1751-7915.13589

Figure Lengend Snippet: Involvement of SodB in copper stress in R. gelatinosus . Western blot analysis of wild‐type ( WT ), ΔcadA, copA − and ΔsodB mutants grown in malate medium supplemented or not (‐) with 100 µM CdCl 2 (Cd 2+ ) or CuSO 4 (Cu 2+ ) (A). Cells were grown by photosynthesis, and total protein extracts from the same amount of cells (0.1 OD 680nm ) were analysed by Western blot using the HRP‐HisProbe. SOD activity assay on non‐denaturing PAGE. Soluble fractions from the ΔcadA, copA , and ΔsodB mutants were separated on 15% PAGE (B). Dose–response curves of the WT , ΔsodB , copA − and ΔsodBcopA − strains. Cells were grown in presence of increasing concentration of CuSO 4 by respiration (C) or under photosynthesis condition (D) for 24 h. Results represent mean ± stdev from 3 independent experiments.

Article Snippet: In the cyanobacterium Synechocystis sp. PCC 6803, high copper treatment also induced the expression of the iron superoxide dismutase sodB (Giner‐Lamia et al. , ).

Techniques: Western Blot, Activity Assay, Concentration Assay

Journal: Microbial Biotechnology

Article Title: Additive effects of metal excess and superoxide, a highly toxic mixture in bacteria

doi: 10.1111/1751-7915.13589

Figure Lengend Snippet: SOD activity was induced in the ATPase mutants of V. cholerae and E. coli. Western blot and SOD in‐gel activity assay of soluble fractions from wild‐type ( WT ) and SodB − strains of V. cholerae (A). SodB activity and expression in V. cholerae were induced in the ATPase‐deficient mutants cadA − and copA − in response to 5 µM CdCl 2 (Cd 2+ ) (B) or 200 µM CuSO 4 (Cu 2+ ) (C) as shown by the in‐gel activity assay and on the Western blots. Effect of excess CuSO 4 on growth of the V. cholerae Δ sodBcopA deletion strain. WT , the single mutants Δ copA, sodB − and the Δ sodBcopA mutant were grown overnight in LB (D) or LB supplemented with 200 µM of CuSO 4 (E), growth was monitored at 600 nm with a Tecan Infinite. Western blot and SOD in‐gel activity assay of soluble fractions from WT, SodA − and SodB − strains of E. coli (F). SodB activity was reduced, while SodA increased in the ATPase mutants zntA − and copA − in response to 20 µM CdCl 2 (Cd 2+ ) (G) or 100 µM CuSO 4 (Cu 2+ ) (H) as shown by the in‐gel activity assays and on the Western blots.

Article Snippet: In the cyanobacterium Synechocystis sp. PCC 6803, high copper treatment also induced the expression of the iron superoxide dismutase sodB (Giner‐Lamia et al. , ).

Techniques: Activity Assay, Western Blot, Expressing, Mutagenesis

Journal: Microbial Biotechnology

Article Title: Additive effects of metal excess and superoxide, a highly toxic mixture in bacteria

doi: 10.1111/1751-7915.13589

Figure Lengend Snippet: SOD activity was induced in the ATPase‐deficient mutants of P. aeruginosa, P. putida and B. subtilis. SodB activity and expression in P. aeruginosa were induced in the ATPase mutants zntA − and copA − in response to 20 µM CdCl 2 or 200 µM CuSO 4 as shown by the in‐gel activity assay and on the Western blot (A). SOD (SodAB) activity and expression in P. putida were induced in the ATPase mutant copA − in response to 50 µM CuSO 4 (B). SodA activity and expression in B. subtilis were induced in the ATPase mutant copA − under 200 µM CuSO 4 stress (C).

Article Snippet: In the cyanobacterium Synechocystis sp. PCC 6803, high copper treatment also induced the expression of the iron superoxide dismutase sodB (Giner‐Lamia et al. , ).

Techniques: Activity Assay, Expressing, Western Blot, Mutagenesis

Journal: Microbial Biotechnology

Article Title: Additive effects of metal excess and superoxide, a highly toxic mixture in bacteria

doi: 10.1111/1751-7915.13589

Figure Lengend Snippet: Interplay between the metal efflux and ROS detoxifying system. Wild‐type (WT) cells induce the expression of the ATPase CadA to prevent Cd 2+ accumulation and related damages and SodB to scavenge superoxide. In the ΔcadA deficient mutant, accumulation of Cd 2+ in the cytosol damage exposed [4Fe‐4S] clusters, cells perceive the situation as an ‘iron‐starvation’ condition and respond by increasing Fe 2+ import presumably to rebuilt [4Fe‐4S]. This led to the induction of the superoxide dismutase SodB in response to iron status. In cells defective in both metal and ROS detoxification systems, the concomitant accumulation of Cd 2+ and ROS leads to metal hypersensitive cells and growth inhibition, because both superoxide and Cd 2+ target enzymes with exposed [4Fe‐4S] clusters. This imbalance and loss of [4Fe‐4S] clusters is no longer tolerable by the bacterium.

Article Snippet: In the cyanobacterium Synechocystis sp. PCC 6803, high copper treatment also induced the expression of the iron superoxide dismutase sodB (Giner‐Lamia et al. , ).

Techniques: Expressing, Mutagenesis, Inhibition