Cysteine homeostasis under inhibition of protein synthesis in Escherichia coli cells

Смирнова, Г. В.; Tyulenev, Aleksey V.; Bezmaternykh, K. V.; Muzyka, Nadezda G.; Ushakov, Vadim Y.; Oktyabrsky, Oleg N.

doi:10.1007/s00726-019-02795-2

Cited by 27 publications

(51 citation statements)

References 51 publications

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“…Accordingly, the concentrations of GSH, GSSG, methionine, and cysteine in E. coli cytosol were calculated to be 3000, 400, 800, and 200 µM, respectively. The concentration for GSH was similar to previous reports, but lower concentrations have been reported for oxidized glutathione, methionine, and cysteine (5, 150, and 20–100 µM, respectively) [ 30 , 32 – 34 ]. No other LMM sulfur species were evident, suggesting that if any were present in cytosol their concentrations must be < 200 µM.…”

Section: Chromatographic Behavior Of Iron and Zinc Standards Reflected The M–l Binding Strength Of The Complexsupporting

confidence: 90%

Low-molecular-mass labile metal pools in Escherichia coli: advances using chromatography and mass spectrometry

Brawley

Lindahl

2021

J Biol Inorg Chem

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Labile low-molecular-mass (LMM) transition metal complexes play essential roles in metal ion trafficking, regulation, and signalling in biological systems, yet their chemical identities remain largely unknown due to their rapid ligand-exchange rates and weak M–L bonds. Here, an Escherichia coli cytosol isolation procedure was developed that was devoid of detergents, strongly coordinating buffers, and EDTA. The interaction of the metal ions from these complexes with a SEC column was minimized by pre-loading the column with 67ZnSO4 and then monitoring 66Zn and other metals by inductively coupled plasma mass spectrometry (ICP-MS) when investigating cytosolic ultrafiltration flow-through-solutions (FTSs). Endogenous cytosolic salts suppressed ESI-MS signals, making the detection of metal complexes difficult. FTSs contained ca. 80 µM Fe, 15 µM Ni, 13 µM Zn, 10 µM Cu, and 1.4 µM Mn (after correcting for dilution during cytosol isolation). FTSs exhibited 2–5 Fe, at least 2 Ni, 2–5 Zn, 2–4 Cu, and at least 2 Mn species with apparent masses between 300 and 5000 Da. Fe(ATP), Fe(GSH), and Zn(GSH) standards were passed through the column to assess their presence in FTS. Major LMM sulfur- and phosphorus-containing species were identified. These included reduced and oxidized glutathione, methionine, cysteine, orthophosphate, and common mono- and di-nucleotides such as ATP, ADP, AMP, and NADH. FTSs from cells grown in media supplemented with one of these metal salts exhibited increased peak intensity for the supplemented metal indicating that the size of the labile metal pools in E. coli is sensitive to the concentration of nutrient metals.

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Section: Chromatographic Behavior Of Iron and Zinc Standards Reflected The M–l Binding Strength Of The Complexsupporting

confidence: 90%

Low-molecular-mass labile metal pools in Escherichia coli: advances using chromatography and mass spectrometry

Brawley

Lindahl

2021

J Biol Inorg Chem

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“…Our studies showed that the transition of E. coli from growth to phosphate starvation is accompanied by H 2 S production, release of free cysteine from the cells into the medium, and an increase in the concentration of intracellular GSH. Previously, similar changes were observed with inhibition of protein synthesis in E. coli in response to the addition of valine or chloramphenicol (Smirnova et al, 2019). These changes can be considered as a non-specific reaction to starvation stress and a way to regulate the level of intracellular cysteine, which is able to potentiate oxidative stress, under conditions of sharp inhibition of protein synthesis.…”

Section: Discussionsupporting

confidence: 65%

“…E. coli has two intracellular thiol redox systems: a glutathione redox system including glutathione (GSH), glutathione reductase (GOR) and glutaredoxins, and a thioredoxin redox system including thioredoxins (TrxA and TrxC) and thioredoxin reductase (TrxB). E. coli cells contain millimolar concentrations of GSH, which participates in maintaining intracellular cysteine homeostasis and serves as the main cytoplasmic redox buffer (Smirnova et al, 2019). The aim of this work was to study the changes in the redox state of glutathione in wild-type E. coli and mutants in various components of thiol redox systems during phosphate starvation.…”

Section: Introductionmentioning

confidence: 99%

Changes in the redox state of Escherichia coli cells during phosphate starvation

Смирнова

Tyulenev

Muzyka

et al. 2020

LFWB

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“…2), whereas a gshB deletion mutant showed an EOP of 61% compared to wild type. GshA and GshB are major components of the glutathione synthesis pathway and the lack of either one results in the absence of intracellular glutathione (43)(44)(45)(46). Glutathione is an important antioxidant and responsible for maintaining a reducing environment in the cell to prevent damage from reactive oxygen species (ROS) such as superoxide and free radicals (47,48).…”

Section: Resultsmentioning

confidence: 99%

The Multidrug Efflux System AcrABZ-TolC Is Essential for Infection of Salmonella Typhimurium by the Flagellum-Dependent Bacteriophage Chi

Esteves

Porwollik

McClelland

et al. 2021

J Virol

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Bacteriophages are the most abundant biological entities in the biosphere. Due to their host specificity and ability to kill bacteria rapidly, bacteriophages have many potential healthcare applications, including therapy against antibiotic-resistant bacteria. Infection by flagellotropic bacteriophages requires a properly rotating bacterial flagellar filament. The flagella-dependent phage χ (Chi) infects serovars of the pathogenic enterobacterium Salmonella enterica. However, cell surface receptors and proteins involved in other stages of χ infection have not been discovered to date. We screened a multi-gene deletion library of S. enterica serovar Typhimurium by spotting mutants on soft agar plates seeded with bacteriophage χ and monitoring their ability to grow and form a swim ring, a characteristic of bacteriophage-resistant motile mutants. Those multi-gene deletion regions identified to be important for χ infectivity were further investigated by characterizing the phenotypes of corresponding single-gene deletion mutants. This way, we identified motile mutants with varying degrees of resistance to χ. Deletions in individual genes encoding the AcrABZ-TolC multi-drug efflux system drastically reduced infection by bacteriophage χ. Furthermore, an acrABtolC triple deletion strain was fully resistant to χ. Infection was severely reduced but not entirely blocked by the deletion of the gene tig encoding the molecular chaperone trigger factor. Finally, deletion in genes encoding enzymes involved in the synthesis of the antioxidants glutathione (GSH) and uric acid resulted in reduced infectivity. Our findings begin to elucidate poorly understood processes involved in later stages of flagellotropic bacteriophage infection and informs research aimed at the use of bacteriophages to combat antibiotic-resistant bacterial infections. IMPORTANCE Antimicrobial resistance is a large concern in the healthcare field. With more multi-drug resistant bacterial pathogens emerging, other techniques for eliminating bacterial infections are being explored. Among these is phage therapy, where combinations of specific phages are used to treat infections. Generally, phages utilize cell appendages and surface receptors for the initial attachment to their host. Phages that are flagellotropic are of particular interest because flagella are often important in bacterial virulence, making resistance to attachment of these phages harder to achieve without reducing virulence. This study discovered the importance of a multi-drug efflux pump for the infection of Salmonella enterica by a flagellotropic phage. In theory, if a bacterial pathogen develops phage resistance by altering expression of the efflux pump then the pathogen would simultaneously become more susceptible to the antibiotic substrates of the pump. Thus, co-administering antibiotics and flagellotropic phage may be a particularly potent antibacterial therapy.

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Cysteine homeostasis under inhibition of protein synthesis in Escherichia coli cells

Cited by 27 publications

References 51 publications

Low-molecular-mass labile metal pools in Escherichia coli: advances using chromatography and mass spectrometry

Low-molecular-mass labile metal pools in Escherichia coli: advances using chromatography and mass spectrometry

Changes in the redox state of Escherichia coli cells during phosphate starvation

The Multidrug Efflux System AcrABZ-TolC Is Essential for Infection of Salmonella Typhimurium by the Flagellum-Dependent Bacteriophage Chi

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