Clustering and Identification of Force Spectra from Native Membranes (original) (raw)
2019, Biophysical Journal
Antimicrobial Resistance (AMR) is an increasing threat to society. Here we have used Atomic Force Microscopy (AFM) to study the bacterial cell wall, which is one of the main antibiotic targets, in Staphylococcus aureus and Bacillus Subtilis. Using high-resolution imaging techniques, the native 3-Dimensional molecular organization of the main component of the cell wall, peptidoglycan, has been resolved for the first time. Direct imaging of the cell wall has allowed previous studies to build models of its 3-Dimensional architecture based on the outer surface of living cells. However, the molecular resolution was far from available at the time. Here we will show that we can visualize single molecules within the living bacterial cell wall. By imaging purified bacteria cell wall (i.e. sacculi), we reveal the organisation of the peptidoglycan architecture on the inner, cytoplasmic side of the cell wall, which is also where synthesis occurs. During the division process, peptidoglycan is mainly inserted in the septum, which now can be directly imaged for the first time showing a heterogeneous structure along the thickness of the cell wall. The architecture observed places hard constraints on the mechanism of cell wall synthesis and cell division in Gram positive bacteria. Using Highresolution Atomic Force Microscopy, we have obtained molecular information that will allow us to start tackling unresolved questions about the bacterial cell wall synthesis machinery and the effect of beta-lactam antibiotics that leads to cell death.
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