Function and X-Ray crystal structure of Escherichia coli YfdE

Mullins, E.A.; Sullivan, Kelly L.; Kappock, T.J.

doi:10.1371/journal.pone.0067901

Cited by 14 publications

(12 citation statements)

References 74 publications

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“…A significant (orders of magnitude) enhancement of the expression for yfdX gene was observed 13 14 compared to other yfdWUVE proteins upon overproduction of evgA in E. coli when quantified using real time PCR. Proteins yfdWUVE are primarily involved in acid tolerance response (ATR) activity 15 . However, yfdX proteins till date remain completely uncharacterized to the best of our knowledge.…”

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Antibiotic binding of STY3178, a yfdX protein from Salmonella Typhi

Saha

Manna

Das

et al. 2016

Sci Rep

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The yfdX family proteins are known for long time to occur in various virulent bacteria including their multidrug resistant (MDR) strains, without any direct assigned function for them. However, yfdX protein along with other proteins involved in acid tolerance response is reported to be up regulated by the multidrug response regulatory system in E. coli. Hence, molecular and functional characterization of this protein is important for understanding of key cellular processes in bacterial cells. Here we study STY3178, a yfdX protein from a MDR strain of typhoid fever causing Salmonella Typhi. Our experimental results indicate that STY3178 is a helical protein existing in a trimeric oligomerization state in solution. We also observe many small antibiotics, like ciprofloxacin, rifampin and ampicillin viably interact with this protein. The dissociation constants from the quenching of steady state fluorescence and isothermal titration calorimetry show that ciprofloxacin binding is stronger than rifampin followed by ampicillin.

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confidence: 99%

Antibiotic binding of STY3178, a yfdX protein from Salmonella Typhi

Saha

Manna

Das

et al. 2016

Sci Rep

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“…The turnover number k cat of Mct with mesaconate and succinyl-CoA (12.2 s -1 ) is comparable to the turnover numbers of other characterized class III CoA-transferases, e.g., acetyl-CoA:oxalate CoA-transferase YfdE from E. coli (22 s -1 ), succinyl-CoA:( S )-malate CoA-transferase from C. aurantiacus (11.4 s -1 ), and succinyl-CoA:3-sulfopropionate CoA-transferase from Variovorax paradoxus (37.7 s -1 ) ( Friedmann et al, 2006 ; Schürmann et al, 2013 ; Mullins et al, 2013 ). All these CoA-transferases use dicarboxylic acids as substrates.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, the known members of class III are highly different in their quaternary structures. Oxalate CoA-transferase, homodimeric acetyl-CoA:oxalate CoA-transferase (YfdE) and butyrobetainyl-CoA:( R )-carnitine CoA-transferase (CaiB) are monomeric ( Baetz and Allison, 1990 ; Stenmark et al, 2004 ; Mullins et al, 2013 ), while benzoylsuccinate CoA-transferase has an α 2 β 2 structure ( Leutwein and Heider, 2001 ), phenyllactate CoA-transferase forms a complex with phenyllactyl-CoA hydratase ( Dickert et al, 2000 ), and succinyl-CoA:L-malate CoA-transferase has an (αβ) 6 structure ( Friedmann et al, 2006 ). Based on size exclusion chromatography data, Mct is a homodimer, like CaiB and YfdE.…”

Section: Discussionmentioning

confidence: 99%

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Succinyl-CoA:Mesaconate CoA-Transferase and Mesaconyl-CoA Hydratase, Enzymes of the Methylaspartate Cycle in Haloarcula hispanica

et al. 2017

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Growth on acetate or other acetyl-CoA-generating substrates as a sole source of carbon requires an anaplerotic pathway for the conversion of acetyl-CoA into cellular building blocks. Haloarchaea (class Halobacteria) possess two different anaplerotic pathways, the classical glyoxylate cycle and the novel methylaspartate cycle. The methylaspartate cycle was discovered in Haloarcula spp. and operates in ∼40% of sequenced haloarchaea. In this cycle, condensation of one molecule of acetyl-CoA with oxaloacetate gives rise to citrate, which is further converted to 2-oxoglutarate and then to glutamate. The following glutamate rearrangement and deamination lead to mesaconate (methylfumarate) that needs to be activated to mesaconyl-C1-CoA and hydrated to β-methylmalyl-CoA. The cleavage of β-methylmalyl-CoA results in the formation of propionyl-CoA and glyoxylate. The carboxylation of propionyl-CoA and the condensation of glyoxylate with another acetyl-CoA molecule give rise to two C4-dicarboxylic acids, thus regenerating the initial acetyl-CoA acceptor and forming malate, its final product. Here we studied two enzymes of the methylaspartate cycle from Haloarcula hispanica, succinyl-CoA:mesaconate CoA-transferase (mesaconate CoA-transferase, Hah_1336) and mesaconyl-CoA hydratase (Hah_1340). Their genes were heterologously expressed in Haloferax volcanii, and the corresponding enzymes were purified and characterized. Mesaconate CoA-transferase was specific for its physiological substrates, mesaconate and succinyl-CoA, and produced only mesaconyl-C1-CoA and no mesaconyl-C4-CoA. Mesaconyl-CoA hydratase had a 3.5-fold bias for the physiological substrate, mesaconyl-C1-CoA, compared to mesaconyl-C4-CoA, and virtually no activity with other tested enoyl-CoA/3-hydroxyacyl-CoA compounds. Our results further prove the functioning of the methylaspartate cycle in haloarchaea and suggest that mesaconate CoA-transferase and mesaconyl-CoA hydratase can be regarded as characteristic enzymes of this cycle.

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“…Exchanged samples were kept at 277 K and used within 12 h. Proteins were quantitated by the method of Bradford using bovine serum albumin as a standard (Bradford, 1976). Gel-filtration analysis was used to determine molecular sizes as described by Mullins et al (2013). Macromolecule-production information is summarized in Table 1.…”

Section: Macromolecule Productionmentioning

confidence: 99%