Breaking antimicrobial resistance by disrupting extracytoplasmic protein folding
- Imperial College London, United Kingdom
- The University of Texas at Austin, United States
- Universidad de Sevilla, Spain
- Brunel University London, United Kingdom
- University of Birmingham, United Kingdom
- Paris-Sud University, France
- University of Neuchatel, Switzerland
Abstract
Antimicrobial resistance in Gram-negative bacteria is one of the greatest threats to global health. New antibacterial strategies are urgently needed, and the development of antibiotic adjuvants that either neutralize resistance proteins or compromise the integrity of the cell envelope is of ever-growing interest. Most available adjuvants are only effective against specific resistance proteins. Here we demonstrate that disruption of cell envelope protein homeostasis simultaneously compromises several classes of resistance determinants. In particular, we find that impairing DsbA-mediated disulfide bond formation incapacitates diverse β-lactamases and destabilizes mobile colistin resistance enzymes. Furthermore, we show that chemical inhibition of DsbA sensitizes multidrug-resistant clinical isolates to existing antibiotics and that the absence of DsbA, in combination with antibiotic treatment, substantially increases the survival of Galleria mellonella larvae infected with multidrug-resistant Pseudomonas aeruginosa. This work lays the foundation for the development of novel antibiotic adjuvants that function as broad-acting resistance breakers.
Data availability
All data generated during this study that support the findings are included in the manuscript or in the Supplementary Information.
Article and author information
Author details
Despoina AI Mavridou
Funding
Medical Research Council (MR/M009505/1)
- Despoina AI Mavridou
Swiss National Science Foundation (PZ00P3_180142)
- Diego Gonzalez
Academy of Medical Sciences (SBF006\1040)
- Ronan R McCarthy
National Institutes of Health (R01AI158753)
- Despoina AI Mavridou
Biotechnology and Biological Sciences Research Council (BB/M02623X/1)
- Jessica MA Blair
Wellcome Trust (105603/Z/14/Z)
- Gerald J Larrouy-Maumus
British Society for Antimicrobial Chemotherapy (BSAC-2018-0095)
- Ronan R McCarthy
Biotechnology and Biological Sciences Research Council (BB/V007823/1)
- Ronan R McCarthy
Swiss National Science Foundation (P300PA_167703)
- Diego Gonzalez
NC3Rs (NC/V001582/1)
- Ronan R McCarthy
Biotechnology and Biological Sciences Research Council (BB/M011178/1)
- Nikol Kaderabkova
Biotechnology and Biological Sciences Research Council (BB/M01116X/1)
- Hannah L Pugh
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2022, Furniss et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Breaking antimicrobial resistance by disrupting extracytoplasmic protein folding
eLife 11:e57974.
https://doi.org/10.7554/eLife.57974
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Teichoic acids (TA) are linear phospho-saccharidic polymers and important constituents of the cell envelope of Gram-positive bacteria, either bound to the peptidoglycan as wall teichoic acids (WTA) or to the membrane as lipoteichoic acids (LTA). The composition of TA varies greatly but the presence of both WTA and LTA is highly conserved, hinting at an underlying fundamental function that is distinct from their specific roles in diverse organisms. We report the observation of a periplasmic space in Streptococcus pneumoniae by cryo-electron microscopy of vitreous sections. The thickness and appearance of this region change upon deletion of genes involved in the attachment of TA, supporting their role in the maintenance of a periplasmic space in Gram-positive bacteria as a possible universal function. Consequences of these mutations were further examined by super-resolved microscopy, following metabolic labeling and fluorophore coupling by click chemistry. This novel labeling method also enabled in-gel analysis of cell fractions. With this approach, we were able to titrate the actual amount of TA per cell and to determine the ratio of WTA to LTA. In addition, we followed the change of TA length during growth phases, and discovered that a mutant devoid of LTA accumulates the membrane-bound polymerized TA precursor.
