Simultaneous determination of phosphonate, phosphate, and diphosphate by capillary electrophoresis using in‐capillary complexation with Mo(VI)

Himeno, Sadayuki; Inazuma, Naoko; Kitano, Eri

doi:10.1002/jssc.200600501

Cited by 17 publications

(12 citation statements)

References 30 publications

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“…The BGE used for electrophoretic experiments was made up of 3 mM sodium molybdate in 200 mM malonate buffer adjusted to pH 3: acetonitrile (ACN) (80∶20; v/v). The determination of PPP i is based on the in-capillary formation of anionic polyoxomolybdate complex in the presence of ACN, as already described for phosphonate, P i and PP i [40]. E. coli supernatants were injected into the CE system after fractionation on a G-25 column.…”

Section: Methodsmentioning

confidence: 99%

High Inorganic Triphosphatase Activities in Bacteria and Mammalian Cells: Identification of the Enzymes Involved

et al. 2012

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BackgroundWe recently characterized a specific inorganic triphosphatase (PPPase) from Nitrosomonas europaea. This enzyme belongs to the CYTH superfamily of proteins. Many bacterial members of this family are annotated as predicted adenylate cyclases, because one of the founding members is CyaB adenylate cyclase from A. hydrophila. The aim of the present study is to determine whether other members of the CYTH protein family also have a PPPase activity, if there are PPPase activities in animal tissues and what enzymes are responsible for these activities.Methodology/Principal FindingsRecombinant enzymes were expressed and purified as GST- or His-tagged fusion proteins and the enzyme activities were determined by measuring the release of inorganic phosphate. We show that the hitherto uncharacterized E. coli CYTH protein ygiF is a specific PPPase, but it contributes only marginally to the total PPPase activity in this organism, where the main enzyme responsible for hydrolysis of inorganic triphosphate (PPPi) is inorganic pyrophosphatase. We further show that CyaB hydrolyzes PPPi but this activity is low compared to its adenylate cyclase activity. Finally we demonstrate a high PPPase activity in mammalian and quail tissue, particularly in the brain. We show that this activity is mainly due to Prune, an exopolyphosphatase overexpressed in metastatic tumors where it promotes cell motility.Conclusions and General SignificanceWe show for the first time that PPPase activities are widespread in bacteria and animals. We identified the enzymes responsible for these activities but we were unable to detect significant amounts of PPPi in E. coli or brain extracts using ion chromatography and capillary electrophoresis. The role of these enzymes may be to hydrolyze PPPi, which could be cytotoxic because of its high affinity for Ca2+, thereby interfering with Ca2+ signaling.

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Section: Methodsmentioning

confidence: 99%

High Inorganic Triphosphatase Activities in Bacteria and Mammalian Cells: Identification of the Enzymes Involved

et al. 2012

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“…The pH and composition of BGE were chosen taking into account the data published in the articles and in accordance with the following criteria: The similarity of the electrophoretic mobilities of the separated ions and main component of BGE. Inertia with respect to the components of the system. Permeability in the selected wavelength range for detection. …”

Section: Resultsmentioning

confidence: 99%

The study of polyoxometalates formation using capillary zone electrophoresis

Zdanov

Shuvaeva

2014

Electrophoresis

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The formation process of polyoxometalates [PMo12 O40 ](3-) and [PMo12 - x Vx O40 ](-3-x) has been studied in aqueous solutions of 0.1 M malonate buffer at pH 2.8-3.0 using CZE. Two different approaches, pre-capillary and in-capillary, were examined and compared. In precapillary mode, the reaction mixture of the reactants and reaction products was injected into the capillary followed by the separation procedure. In in-capillary mode, the sequential input of the reagents and running electrolyte into the capillary and the species separation occurs simultaneously. The optimal parameters of in-capillary separation were established as functions of applied voltage and the length of the intermediate buffer zone between the reagents in the capillary. As a result the best-compromise conditions for the separation of the mixtures containing the reactants, intermediates, and reaction products, in order to achieve the best efficiency, symmetry, and peak areas, were achieved at -18 kV and the input parameter of 900 mbar·s. It was also shown that in-capillary mode is more informative than pre-capillary mode for studying the complex compound formation process.

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“…In the selection of an electrolyte composition for the separation of polyoxo metalates by CZE, we were guided by the data [22,24]. Taking into account the stability of complex ions, preference was given to electrolytes based on organic acids.…”

Section: Methodsmentioning

confidence: 99%

“…It should be noted that the CZE data strongly differ from the IP RP HPLC data obtained in the study of similar systems [21]. Few examples of using capillary electrophoresis in the chemistry of polyoxoanions, as a rule, were associated with the determination of trace elements as heteropolyoxoanions (molybdates or tungstates) absorbing in the UV region [21][22][23][24].…”

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confidence: 96%

A study of complex phosphovanadomolybdates [PV x Mo12−x O40]−(3+x) by reversed-phase HPLC and capillary zone electrophoresis

Zhdanov

Shuvaeva

2015

J Anal Chem

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A fundamental possibility of using ion pair reversed phased HPLC and capillary zone electro phoresis (CZE) for the study of polyoxometalate (POM) solutions has been demonstrated on an example of phosphovanadomolybdates of the composition [PV x Mo 12 -x O 40 ] -(3 + x), where x = 1-3 and the possibility of the separation of the products of their synthesis has been shown. It has been found that the modification of the inner surface of a capillary by alkyl groups C 1 and C 8 results in an increase in the resolution of analytical signals in CZE. It has been shown that the results obtained by HPLC and CZE are in a good agreement and can be confirmed by the data of 51 V NMR spectrometry.

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Simultaneous determination of phosphonate, phosphate, and diphosphate by capillary electrophoresis using in‐capillary complexation with Mo(VI)

Cited by 17 publications

References 30 publications

High Inorganic Triphosphatase Activities in Bacteria and Mammalian Cells: Identification of the Enzymes Involved

High Inorganic Triphosphatase Activities in Bacteria and Mammalian Cells: Identification of the Enzymes Involved

The study of polyoxometalates formation using capillary zone electrophoresis

A study of complex phosphovanadomolybdates [PV x Mo12−x O40]−(3+x) by reversed-phase HPLC and capillary zone electrophoresis

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