Altered Envelope Structure and Nanomechanical Properties of a C-Terminal Protease A-Deficient Rhizobium leguminosarum
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains, Plasmids, and Media
2.2. Isolation of Peptidoglycan
2.3. Muropeptide Structural Analysis
2.4. Transmission Electron Microscopy (TEM) Imaging
2.5. AFM Imaging and Force Data Acquisition
2.6. AFM Creep Experiments
3. Results
3.1. Cell Envelope Ultrastructure
3.2. Muropeptide Composition
3.3. R. leguminosarum Cell Envelope Elasticity and Spring Constant
3.4. Viscoelasticity of the Cell Envelope
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Peak | Proposed Structure | Observed m/z | Calculated [M+Na]+ | % of All Peaks | |
---|---|---|---|---|---|
Wild Type | ctpA Mutant | ||||
2 | GlcNAc-MurNAc-Ala-Glu-meso-DAP | 893.4 | 893.8 | 2.0 | 2.7 |
3 | GlcNAc-MurNAc-Ala-Glu-meso-DAP-Gly | 950.4 | 950.9 | 1.6 | 1.6 |
6 | GlcNAc-MurNAc-Ala-Glu-meso-DAP-Ala | 964.5 | 964.9 | 6.2 | 8.3 |
10 | GlcNAc-MurNAc-Ala-Glu-meso-DAP-Ala-meso-DAP-Glu-Ala-MurNAc-GlcNAc | 1816.9 | 1816.8 | 1.3 | 2.4 |
11 | MurNAc-Ala-Glu-meso-DAP-Ala-meso-DAP-Glu-Ala-MurNAc | 1408.7 | 1408.4 | 1.1 | 2.2 |
12 | GlcNAc-MurNAc-Ala-Glu-meso-DAP-Ala-Ala-meso-DAP-Glu-Ala-MurNAc-GlcNAc | 1888.0 | 1887.9 | 2.9 | 3.0 |
Force (nN) | k1 (N/m) | k2 (N/m) | η2 (N•s/m) | η1 (N•s/m) | ||||
---|---|---|---|---|---|---|---|---|
wt | ctpA | wt | ctpA | wt | ctpA | wt | ctpA | |
2 | 0.52 ± 0.37 n = 22 | 0.45 ± 0.22 n = 21 | 0.22 ± 0.12 n = 22 | 0.18 ± 0.06 n = 21 | 0.14 ± 0.12 n = 22 | 0.13 ± 0.06 n = 21 | 5.27 ± 4.38 n = 22 | 5.04 ± 4.24 n = 21 |
4 | 0.89 ± 0.48 n = 26 | 0.758 ± 0.46 n = 23 | 0.31 ± 0.13 n = 26 | 0.30 ± 0.13 n = 23 | 0.23 ± 0.15 n = 26 | 0.23 ± 0.10 n = 23 | 8.22 ± 6.26 n = 26 | 7.51 ± 5.58 n = 23 |
6 | 0.97 ± 0.56 * n = 18 | 1.39 ± 0.78 * n = 23 | 0.37 ± 0.12 n = 18 | 0.41 ± 0.16 n = 23 | 0.27 ± 0.16 n = 18 | 0.33 ± 0.23 n = 23 | 9.95 ± 5.66 * n = 18 | 7.04 ± 4.50 * n = 23 |
10 | 1.56 ± 0.59 n = 21 | 1.71 ± 0.92 n = 27 | 0.57 ± 0.15 * n = 21 | 0.46 ± 0.21 * n = 27 | 0.52 ± 0.36 n = 21 | 0.48 ± 0.21 n = 27 | 12.5 ± 5.97 † n = 21 | 6.78 ± 3.87 † n = 27 |
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Jun, D.; Idem, U.; Dahms, T.E.S. Altered Envelope Structure and Nanomechanical Properties of a C-Terminal Protease A-Deficient Rhizobium leguminosarum. Microorganisms 2020, 8, 1421. https://doi.org/10.3390/microorganisms8091421
Jun D, Idem U, Dahms TES. Altered Envelope Structure and Nanomechanical Properties of a C-Terminal Protease A-Deficient Rhizobium leguminosarum. Microorganisms. 2020; 8(9):1421. https://doi.org/10.3390/microorganisms8091421
Chicago/Turabian StyleJun, Dong, Ubong Idem, and Tanya E. S. Dahms. 2020. "Altered Envelope Structure and Nanomechanical Properties of a C-Terminal Protease A-Deficient Rhizobium leguminosarum" Microorganisms 8, no. 9: 1421. https://doi.org/10.3390/microorganisms8091421