Molecular Basis of AmpC β-Lactamase Induction by Avibactam in Pseudomonas aeruginosa: PBP Occupancy, Live Cell Binding Dynamics and Impact on Resistant Clinical Isolates Harboring PDC-X Variants
Abstract
:1. Introduction
2. Results and Discussion
2.1. Molecular Basis of Avibactam AmpC Induction
2.2. Avibactam Target Binding in Live P. aeruginosa Cells
2.3. qRT-PCR-Based ampC Induction Assays
3. Methods
3.1. Bacterial Strains and Antibiotics
3.2. Intact Cells’ Time-Course of PBP-Binding Assay
3.3. Determination of PBP-Binding Affinity (IC50)
3.4. qRT-PCR Quantification of AmpC Induction
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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P. aeruginosa PAO1 PBP a | Mean IC50 ± SD (µg/mL) b | ||
---|---|---|---|
Imipenem | Cefoxitin | Avibactam | |
1a | 0.2 ± 0.06 | 7.8 ± 0.5 | >32 |
1b | 0.1 ± 0.03 | >32 | 3.6 ± 0.1 |
2 | 0.05 ± 0.02 | 14.6 ± 2.2 | 4.2 ± 1.6 |
3 | 0.3 ± 0.07 | 10.7 ± 1.7 | >32 |
4 | 0.1 ± 0.04 | 1.5 ± 0.5 | 3.1 ± 1.3 |
5/6 | 0.5 ± 0.2 | 9 ± 2.4 | 2.2 ± 0.7 |
P. aeruginosa Strain a | ampC Expression (min) b | ||
---|---|---|---|
0 | 30 | 180 | |
PAO1 | 1 | 2.6 ± 0.5 | 5.8 ± 0.1 |
PAΔdacB | 110 ± 0.8 | 228 ± 117 | 1100 ± 243 |
PAΔdacBdacCpbpG | 7500 ± 3800 | 27,000 ± 9400 | 58,000 ± 26,000 |
101-E5 | 554 ± 273 | 466 ± 272 | 2400 ± 1100 |
103-H8 | 391 ± 345 | 317 ± 21.2 | 5400 ± 2900 |
109-E9 | 414 ± 122 | 496 ± 213 | 1700 ± 382 |
110-G8 | 306 ± 31.2 | 418 ± 183 | 3500 ± 1600 |
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López-Argüello, S.; Montaner, M.; Oliver, A.; Moya, B. Molecular Basis of AmpC β-Lactamase Induction by Avibactam in Pseudomonas aeruginosa: PBP Occupancy, Live Cell Binding Dynamics and Impact on Resistant Clinical Isolates Harboring PDC-X Variants. Int. J. Mol. Sci. 2021, 22, 3051. https://doi.org/10.3390/ijms22063051
López-Argüello S, Montaner M, Oliver A, Moya B. Molecular Basis of AmpC β-Lactamase Induction by Avibactam in Pseudomonas aeruginosa: PBP Occupancy, Live Cell Binding Dynamics and Impact on Resistant Clinical Isolates Harboring PDC-X Variants. International Journal of Molecular Sciences. 2021; 22(6):3051. https://doi.org/10.3390/ijms22063051
Chicago/Turabian StyleLópez-Argüello, Silvia, María Montaner, Antonio Oliver, and Bartolome Moya. 2021. "Molecular Basis of AmpC β-Lactamase Induction by Avibactam in Pseudomonas aeruginosa: PBP Occupancy, Live Cell Binding Dynamics and Impact on Resistant Clinical Isolates Harboring PDC-X Variants" International Journal of Molecular Sciences 22, no. 6: 3051. https://doi.org/10.3390/ijms22063051