Kristina M Smith, Yigong Bu, Hiroaki Suga Chemistry & Biology

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Induction and Inhibition of Pseudomonas aeruginosa Quorum Sensing by Synthetic Autoinducer Analogs Kristina M Smith, Yigong Bu, Hiroaki Suga Chemistry & Biology Volume 10, Issue 1, Pages 81-89 (January 2003) DOI: 10.1016/S1074-5521(03)00002-4

Figure 1 Chemical Structures of Autoinducers and Analogs AI1, 3-oxo-C12HSL; AI2, C4 HSL; 1, 3-oxo-C12-(2-aminocyclopentanone); 2, 3-oxo-C12-(2-aminocyclopentanol); 3, 3-oxo-C12-(2-aminocyclohexanone); 4, 3-oxo-C12-(2-aminocyclohexanol); 5, C4-(2-aminocyclopentanone); 6, C4-(2-aminocyclopentanol); 7, C4-(2-aminocyclohexanone); 8, C4-(2-aminocyclohexanol). Chemistry & Biology 2003 10, 81-89DOI: (10.1016/S1074-5521(03)00002-4)

Figure 2 Reporter Gene Induction by Autoinducers and Their Analogs (A) Molecular Imager scan of 96-well plate containing PAO-JP2 (plasI-LVAgfp) in the presence of 1, 100, and 400 μM of the designated 3-oxo-C12 compound. (B) Quantification of results of 3-oxo-C13 compounds. Cont, negative control of untreated cell (not shown in [A]). Average fluorescence of four replicate wells was corrected for cell density by dividing by OD600 of cell culture. The standard deviation was derived from three independent experiments shown in (A). (C) Molecular Imager scan of 96-well plated containing PAO-JP2 (prhlI-LVAgfp) in the presence of 1 μM AI1 and 10, 100, and 400 μM designated C4 compound. (D) Quantification of results of C4 compounds. Data analysis was the same as (B). Chemistry & Biology 2003 10, 81-89DOI: (10.1016/S1074-5521(03)00002-4)

Figure 3 Antagonist Assays (A) GFP expression by PAO-JP2 (plasI-LVAgfp) in the presence of 1 μM AI1 alone (control, black bar) or 1 μM AI1 plus 2 or 3. (B) GFP expression by PAO-JP2 (prhlI-LVAgfp) in the presence of 1 μM AI1 and 10 μM AI2 (control, black bar) or 1 μM AI1 and 10 μM AI2 plus 2 or 3. Chemistry & Biology 2003 10, 81-89DOI: (10.1016/S1074-5521(03)00002-4)

Figure 4 Virulence Factor Assays (A) Pyocyanin expression by PAO-JP2. −, negative control; +, positive control, 25 μM each of AI1 and AI2; 2, 25 μM each of AI1 and AI2 in the presence of 100 μM 2; 3, 25 μM each of AI1 and AI2 in the presence of 100 μM 3. (B) Quantification of pyocyanin expression by PAO-JP2 in the presence of various concentrations of 2 or 3. −, negative control containing no AIs (black bar); +, positive control containing 25 μM each of AI1 and AI2 (purple bar); 2, 25 μM each of AI1 and AI2 in the presence of 2; 3, 25 μM each of AI1 and AI2 in the presence of 3. (C) Quantification of pyocyanin expression of wild-type strain PAO1. The conditions are the same as (B), except for control (purple bar) where no AIs were added. (D) Elastase B activity produced by PAO-JP2 in the absence (negative control, black bar) or presence of 5 μM AI1 and 10 μM AI2 (positive control, purple bar) or 5 μM AI1 and 10 μM AI2 plus 2 or 3. (E) Elastase B activity produced by PAO1 control (purple bar) or in the presence of 2 or 3. Chemistry & Biology 2003 10, 81-89DOI: (10.1016/S1074-5521(03)00002-4)

Figure 5 Static Biofilm Assay (A) PAO-JP2 (pTdK-GFP) biofilm development in the presence of designated compounds. (B) PAO-1 (pTdK-GFP) biofilm development in the absence (negative control) or presence of 50 μM 3. Chemistry & Biology 2003 10, 81-89DOI: (10.1016/S1074-5521(03)00002-4)