quadtec uv-vis detector Search Results


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(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) <t>UV-visible</t> absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.
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(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) <t>UV-visible</t> absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.
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(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) <t>UV-visible</t> absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.
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Akzo Nobel kromasil eternity-5-phenylhexyl column
(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) <t>UV-visible</t> absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.
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(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) <t>UV-visible</t> absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.
Kromasil Eternity 5 Phenylhexyl Column, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) <t>UV-visible</t> absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.
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(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) <t>UV-visible</t> absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.
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Image Search Results


(A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) UV-visible absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.

Journal: The Journal of Clinical Investigation

Article Title: Enterobacteria secrete an inhibitor of Pseudomonas virulence during clinical bacteriuria

doi: 10.1172/JCI92464

Figure Lengend Snippet: (A) Multiple chemical characterizations identify escherichelin as HPTT-COOH, a compound shown to inhibit pyochelin-mediated iron uptake in Pseudomonas. The positions of 13C salicylate incorporation are indicated in green, while MS/MS neutral losses are indicated as dashed lines. (B) Mass spectra at the retention time for escherichelin from WT UTI89, UTI89ΔybtS, and 13C6-SA–supplemented UTI89ΔybtS culture supernatants revealed that 6 carbons in escherichelin are derived from salicylate. (C) MS/MS product ion spectrum of escherichelin. The 46-Da neutral loss indicates the presence of a carboxylic acid group, and the 87- and 104-Da losses correspond to fragmentation within the thiazoline ring. (D) UV-visible absorption spectra of purified escherichelin incubated with equimolar FeCl3 or CuSO4. (E and F) DFT simulations of escherichelin bound to Fe(III) in a 1:1 (E) and 2:1 (F) complex. The simulations predict stable Fe(III)-escherichelin complexes with hexacoordinate octahedral geometry. (G) UTI89 growth curve in the presence or absence of a ferric ion chelator (0.5 mM EDDHA) and/or ferric ion supplementation (1 μM FeCl3). Growth was monitored by OD at 600 nm (OD600). Data represent the mean of 3, with SD plotted. (H) PW5011 (pvdA-E02::ISlacZ/hah), a P. aeruginosa strain MPAO1 transposon mutant (55, 56) expressing pyochelin as its sole siderophore, was grown in succinate medium for 20 hours in the presence of increasing escherichelin concentrations relative to vehicle control. Bacterial growth was quantified by enumerating CFU/ml and is expressed as a percentage of the vehicle control. Data represent the mean of 3, with the SEM plotted. *P < 0.01 and **P < 0.001, by 1-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: The eluate was dried down with a lyophilizer (Labconco), resuspended in 20% methanol, and further purified on a Bio-Rad BioLogic DuoFlow 10 system equipped with a QuadTec UV-Vis detector and a Kromasil Eternity-5-phenylhexyl column (AkzoNobel).

Techniques: Tandem Mass Spectroscopy, Derivative Assay, Purification, Incubation, Mutagenesis, Expressing