Journal: Biomolecules

Article Title: Identification of Small Molecules Blocking the Pseudomonas aeruginosa Type III Secretion System Protein PcrV

doi: 10.3390/biom11010055

Figure Lengend Snippet: ( A ) Surface plasmon resonance data for compound H1 binding to PcrV at 400, 200 (duplicate), 100, 50, 25, 12.5, 6.25, and 0 µM. ( B ) Equilibrium analysis of compound H1 from binding levels at 45-55 s in A : K D was determined to be 98.3 µM. R max was 71.3 RU, and Chi 2 was 10.0 RU. ( C ) Infection assay with dose-response analysis of compound H1 that protects macrophages from T3SS-mediated P. aeruginosa toxicity (wt): uninfected cells and infection with a pcrV mutant were used as controls, and standard deviation was calculated by Gaussian approximation. ( D ) 1 H NMR relaxation-edited binding experiment showing the aromatic region for compound H1 at 100 µM with (bottom trace) and without 10 µM PcrV (upper trace): equilibrium analysis was performed in the ProteOn Manager software TM (Bio-Rad Laboratories Inc., Hercules, CA, USA). ( E ) Bacterial growth assay where P. aeruginosa was grown in the presence of compound H1 at 200, 100, 50, and 25 µM and OD600 was measured regularly for 8 h: each value represents the mean value of a triplicate; to the control wells, 1% DMSO was added (DMSO). The panels show one representative experiment.

Article Snippet: The plates were sealed (ProteOn Microplate sealing film, Bio-Rad Laboratories Inc., Hercules, CA, USA) when the plate was prepared.

Techniques: SPR Assay, Binding Assay, Infection, Mutagenesis, Standard Deviation, Software, Growth Assay, Control