Article Figures & Data
Figures
- FIG 1
Bactericidal effects of SMAP-29, KZ144, Art-085, and Art-175 against P. aeruginosa PAO1. A cell suspension was treated with equimolar amounts of SMAP-29, KZ144, Art-085, and Art-175 (3 μM) in the absence and presence of 0.125 or 0.5 mM EDTA. Bacterial reduction after 1 h is expressed in log10 units relative to the untreated control. Mean values (± standard deviation [SD]) are shown. An asterisk indicates when the detection limit (5.15 log units) was reached.
- FIG 2
Rapid mode of action of Art-175. (A) P. aeruginosa PAO1 is rapidly lysed upon contact with Art-175 (25× MIC). Shortly after adding Art-175, a hole is the cell wall is punctured and the cytoplasmic membrane increasingly bulges until the cell undergoes abrupt lysis (see also Movie S1 in the supplemental material). (B) A 30-min time course shows that P. aeruginosa PAO1 treated with 25× MIC Art-175 is reduced by approximately 3 log units in the first 5 min, increasing to 4 log units after 30 min. Mean values (± SD) are shown.
- FIG 3
Resistance development against Art-175 and ciprofloxacin. A laboratory strain of clinical origin (P. aeruginosa PAO1) (A), an environmental strain (P. aeruginosa Br257) (B), and a multidrug-resistant strain (P. aeruginosa Br776) (C) were serially exposed to subinhibitory concentrations to select for decreased susceptibility. Over 20 passages, the MIC of Art-175 (◊) increased 2-fold, whereas the MIC of ciprofloxacin (☐) increased 16- (PAO1), 64- (Br257), or 4-fold (Br776).
- FIG 4
Persister killing by Art-175. The bactericidal effects of Art-175 at 10× and 30× MIC on isolated persister fractions of P. aeruginosa PA14 (light gray) and PA1255 (dark gray) are compared to those of ciprofloxacin at 10× MIC and 30× MIC, in both the absence and the presence of 0.5 mM EDTA. Controls include 0.5 mM EDTA and ofloxacin (10× MIC and 30× MIC). Mean values (± standard error of the mean [SEM]) are shown for at least three independent replicates.
Tables
- TABLE 1
MICs of Art-085 and Art-175a
EDTA concn (mM) or parameter MIC (μg/ml) for: Art-085 Art-175 0 18 10 0.125 20 10 0.25 16 6 0.5 10 4 1 6 4 2 4 2 4 4 2 MIC50b 8 4 MIC90 15 10 ↵a Art-175 (10 μg/ml) has an improved inhibitory activity against P. aeruginosa PAO1 compared to that of Art-085 (18 μg/ml). From 0.25 mM EDTA, MICs further decrease to 4 and 2 μg/ml for Art-085 and Art-175, respectively.
↵b The MICs at which growth of 50% (MIC50) or 90% (MIC90) of the isolates are inhibited, are shown.
Supplemental material
Files in this Data Supplement:
- Supplemental file 1 -
Supplemental Figures S1 to S3 and Tables S1 to S3.
PDF, 658K
- Supplemental file 2
-
Supplemental Movie S1: time-lapse series during exposure of P. aeruginosa to Art-175 (25× MIC, 0.5 mM EDTA). Recording started ~1 min after addition of the protein (scale bar = 4 µm; 5 s per frame; total duration, 21 min 5 s).
AVI, 8.0M
- Supplemental file 3
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Supplemental Movie S2: time-lapse series during exposure of P. aeruginosa to Art-175 (30× MIC, 0.5 mM EDTA). Recording started ~1 min after addition of the protein (scale bar = 4 µm; 5 s per frame; total duration, 5 min 25 s).
AVI, 2.5M
- Supplemental file 4
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Supplemental Movie S3: time-lapse series during exposure of P. aeruginosa to SMAP-29 (25× MIC, 0.5 mM EDTA). Recording started ~1 min after addition of the protein (scale bar = 4 µm; 2 min per frame; total duration, 3 h 48 min).
AVI, 3.8M
- Supplemental file 1 -
