Membrane Transporters Involved in the Antimicrobial Activities of Pyrithione in Escherichia coli
Abstract
:1. Introduction
2. Results
2.1. Screening Membrane Protein Genes of the E. coli Keio Collection: Identification of Membrane Proteins Potentially Involved in the Pharmacological Actions of Pyrithione
2.2. Growth Kinetics of Knockout and Overexpression Strains for metQ, fepC, copA, and zntA Genes at Variable Concentrations of Pyrithione Expanded on the Observations from the Initial Knockout Library Screening
2.3. Effects of the Gene Knockouts on the Minimum Inhibitory Concentrations of Pyrithione in the Presence and Absence of Relevant Metals
2.4. Levels of Cell-Associated Metals in the Absence of Membrane Transporters CopA, ZntA, FepC, and MetQ
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Strains and Culture
5.2. Growth Profiling
5.3. Measurement of Cell Associated Copper
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Media | CuCl2 (µM) | IC50 (µM) |
---|---|---|
Lysogeny Broth (LB) | 0 | 6.05 ± 1.02 |
5 | 5.22 ± 0.95 | |
10 | 0.78 ± 0.25 | |
20 | 0.81 ± 0.28 |
Strain (KO) | Growth Rate Ratio | Gene Annotation | |
---|---|---|---|
Mean | SD | ||
metQ | 2.41 | 0.92 | L-methionine/D-methionine ABC transporter membrane anchored binding protein |
fepC | 2.12 | 0.90 | ferric enterobactin ABC transporter ATP binding subunit |
chbA | 1.75 | 0.70 | N,N’-diacetylchitobiose-specific PTS enzyme IIA component |
araJ | 1.67 | 0.64 | putative transport protein AraJ |
phoR | 1.66 | 0.46 | sensor histidine kinase |
lldP | 1.59 | 0.41 | lactate/glycolate:H+ symporter LldP |
potB | 1.59 | 0.46 | spermidine preferential ABC transporter membrane subunit PotB |
glvB | 1.54 | 0.45 | putative PTS enzyme II component GlvB |
hofC | 1.54 | 0.38 | inner membrane protein HofC |
mdtC | 1.52 | 0.31 | multidrug efflux pump RND permease subunit MdtC |
WT | 1.04 | 0.11 | E. coli BW25113 (Keio collection reference strain) |
tatC | 0.70 | 0.11 | twin arginine protein translocation system-TatC protein |
yjeP | 0.70 | 0.13 | mini conductance mechanosensitive channel MscM |
fucP | 0.69 | 0.19 | L-fucose:H+ symporter |
ybiT | 0.69 | 0.14 | putative ATP-binding protein YbiT |
yheS | 0.60 | 0.20 | putative ATP-binding protein YheS |
malF | 0.54 | 0.10 | maltose ABC transporter membrane subunit MalF |
xylE | 0.52 | 0.16 | D-xylose:H+ symporter |
atpA | 0.44 | 0.12 | ATP synthase F1 complex subunit α |
copA | 0.44 | 0.12 | Cu+ exporting P-type ATPase (copA;ybaR;atcU) |
zntA | 0.09 | 0.03 | Zn2+/Cd2+/Pb2+ exporting P-type ATPase |
Conditions | BW25113 | metQ | fepC | copA | zntA |
---|---|---|---|---|---|
PT alone | 17.5 | 17.5 | 17.5 | 8.8 | 2.2 |
PT and 10 µM Cu | 17.5 | 17.5 | 17.5 | 2.2 | 8.8 |
PT and 10 µM Fe | 17.5 | 17.5 | 17.5 | 8.8 | 2.2 |
PT and 10 µM Zn | 17.5 | 8.8 | 17.5 | 8.8 | 1.1 |
PT and 1.92 mM BCS | >140 | >140 | >140 | >140 | 140 |
PT and 1.92 mM BCS and 10 µM Zn | >140 | >140 | 140 | >140 | 17.5 |
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Salcedo-Sora, J.E.; Robison, A.T.R.; Zaengle-Barone, J.; Franz, K.J.; Kell, D.B. Membrane Transporters Involved in the Antimicrobial Activities of Pyrithione in Escherichia coli. Molecules 2021, 26, 5826. https://doi.org/10.3390/molecules26195826
Salcedo-Sora JE, Robison ATR, Zaengle-Barone J, Franz KJ, Kell DB. Membrane Transporters Involved in the Antimicrobial Activities of Pyrithione in Escherichia coli. Molecules. 2021; 26(19):5826. https://doi.org/10.3390/molecules26195826
Chicago/Turabian StyleSalcedo-Sora, Jesus Enrique, Amy T. R. Robison, Jacqueline Zaengle-Barone, Katherine J. Franz, and Douglas B. Kell. 2021. "Membrane Transporters Involved in the Antimicrobial Activities of Pyrithione in Escherichia coli" Molecules 26, no. 19: 5826. https://doi.org/10.3390/molecules26195826