Oxygen‐insensitive nitroreductase <i>E. coli</i><scp>NfsA</scp>, but not <scp>NfsB</scp>, is inhibited by fumarate

Day, Martin A.; Jarrom, David; Rajah, Navina; Searle, Peter F.; Hyde, Eva I.; White, Scott A.

doi:10.1002/prot.26451

Cited by 3 publications

(1 citation statement)

References 47 publications

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“…The supernatant was loaded onto Ni-NTA resin (Sangon, Shanghai, China), washed with washing buffer (20 mM Tris, 200 mM NaCl, 40 mM imidazole, pH 7.0), and then eluted with an elution buffer (20 mM Tris, 200 mM NaCl, 250 mM imidazole, pH 7.0). The eluted NfsB solution was dialyzed against 4 L of 25 mM HEPES buffer (pH 7.6) overnight, and then quantified via UV-VIS absorption, [34] aliquoted, and stored at À 80 °C.…”

Section: Methodsmentioning

confidence: 99%

Hemicyanine‐Based Highly Water‐Soluble Probe for Extracellular Nitroreductase

Jiang,

Xu,

Zhong

et al. 2024

ChemBioChem

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Nitroreductase (NTR) has long been a target of interest for its important role involved in the nitro compounds metabolism. Various probes have been reported for NTR analysis, but rarely able to distinguish the extracellular NTR from intracellular ones. Herein we reported a new NTR sensor, HCyS‐NO2, which was a hemicyanine molecule with one nitro and two sulfo groups attached. The nitro group acted as the reporting group to respond NTR reduction. Direct linkage of nitro group into the hemicyanine π conjugate system facilitated the intramolecular electron transfer (IET) process and thus quenched the fluorescence of hemicyanine core. Upon reduction with NTR, the nitro group was rapidly converted into the hydroxylamino and then the amino group, eliminating IET process and thus restoring the fluorescence. The sulfo groups installed significantly increased the hydrophilicity of the molecule, and introduced negative charges at physiological pH, preventing the diffusion into bacteria. Both gram‐negative and gram‐positive bacteria were able to turn on the fluorescence of HCyS‐NO2, without detectable diffusion into cells, providing a useful tool to probe the extracellular reduction process.

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Section: Methodsmentioning

confidence: 99%

Hemicyanine‐Based Highly Water‐Soluble Probe for Extracellular Nitroreductase

Jiang,

Xu,

Zhong

et al. 2024

ChemBioChem

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Microbiota and other detrimental metabolites in colorectal cancer

González,

Fullaondo,

Odriozola

et al. 2024

Advances in Genetics

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Antibiotic resistance begets more resistance: chromosomal resistance mutations mitigate fitness costs conferred by multi-resistant clinical plasmids

Nair,

Andersson,

Warsi

2024

Microbiol Spectr

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Plasmids are the primary vectors of horizontal transfer of antibiotic resistance genes among bacteria. Previous studies have shown that the spread and maintenance of plasmids among bacterial populations depend on the genetic makeup of both the plasmid and the host bacterium. Antibiotic resistance can also be acquired through mutations in the bacterial chromosome, which not only confer resistance but also result in changes in bacterial physiology and typically a reduction in fitness. However, it is unclear whether chromosomal resistance mutations affect the interaction between plasmids and the host bacteria. To address this question, we introduced 13 clinical plasmids into a susceptible Escherichia coli strain and three different congenic mutants that were resistant to nitrofurantoin (Δ nfsAB ), ciprofloxacin ( gyrA, S83L), and streptomycin ( rpsL, K42N) and determined how the plasmids affected the exponential growth rates of the host in glucose minimal media. We find that though plasmids confer costs on the susceptible strains, those costs are fully mitigated in the three resistant mutants. In several cases, this results in a competitive advantage of the resistant strains over the susceptible strain when both carry the same plasmid and are grown in the absence of antibiotics. Our results suggest that bacteria carrying chromosomal mutations for antibiotic resistance could be a better reservoir for resistance plasmids, thereby driving the evolution of multi-drug resistance. IMPORTANCE Plasmids have led to the rampant spread of antibiotic resistance genes globally. Plasmids often carry antibiotic resistance genes and other genes needed for its maintenance and spread, which typically confer a fitness cost on the host cell observed as a reduced growth rate. Resistance is also acquired via chromosomal mutations, and similar to plasmids they also reduce bacterial fitness. However, we do not know whether resistance mutations affect the bacterial ability to carry plasmids. Here, we introduced 13 multi-resistant clinical plasmids into a susceptible and three different resistant E. coli strains and found that most of these plasmids do confer fitness cost on susceptible cells, but these costs disappear in the resistant strains which often lead to fitness advantage for the resistant strains in the absence of antibiotic selection. Our results imply that already resistant bacteria are a more favorable reservoir for multi-resistant plasmids, promoting the ascendance of multi-resistant bacteria.

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Oxygen‐insensitive nitroreductase E. coliNfsA, but not NfsB, is inhibited by fumarate

Cited by 3 publications

References 47 publications

Hemicyanine‐Based Highly Water‐Soluble Probe for Extracellular Nitroreductase

Hemicyanine‐Based Highly Water‐Soluble Probe for Extracellular Nitroreductase

Microbiota and other detrimental metabolites in colorectal cancer

Antibiotic resistance begets more resistance: chromosomal resistance mutations mitigate fitness costs conferred by multi-resistant clinical plasmids

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