Development of a capillary electrophoresis‐mass spectrometry method using polymer capillaries for metabolomic analysis of yeast

Tanaka, Yoshihide; Higashi, Tetsuji; Rakwal, Randeep; Wakida, Shin‐ichi; Iwahashi, Hitoshi

doi:10.1002/elps.200700466

Cited by 29 publications

(14 citation statements)

References 31 publications

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“…27,38 For many biological samples, proteins and peptides adsorbed onto the silica surface during separation reduced the repeatability of measurement. Different coatings have been applied to the inner surface of the capillary to improve the repeatability; 27,39-42 however, they reduced the quality of the nanospray in our experiments. Generally, a number of parameters are associated with a stable spray and a successful CE-MS measurement, including the emitter geometry, interface design, buffer composition, SL flow rate, spray voltage and cleanliness of the interface.…”

Section: Resultsmentioning

confidence: 99%

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Liu

Aerts

Rubakhin

et al. 2014

Analyst

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Analytical technologies that enable investigations at the single cell level facilitate a range of studies; here a lab-fabricated capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) platform was used to analyze anionic metabolites from individual Aplysia californica neurons. The system employs a customized coaxial sheath-flow nanospray interface connected to a separation capillary, with the sheath liquid and separation buffer optimized to ensure a stable spray. The method provided good repeatability of separation and reliable detection sensitivity for 16 mono-, di- and triphosphate nucleosides. For a range of anionic analytes, including cyclic adenosine monophosphate (cAMP), adenosine diphosphate (ADP) and adenosine triphosphate (ATP), the detection limits were in the low nanomolar range (<22 nM). A large Aplysia R2 neuron was used to demonstrate the ability of CE-ESI-MS to quantitatively characterize anionic metabolites within individual cells, with 15 nucleotides and derivatives detected. Following the method validation process, we probed smaller, 60-μm diameter Aplysia sensory neurons where sample stacking was used as a simple on-line analyte preconcentration approach. The calculated energy balance ([ATP] + 0.5 × [ADP])/([AMP] + [ADP] + [ATP]) of these cells was comparable with the value obtained from bulk samples.

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Section: Resultsmentioning

confidence: 99%

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Liu

Aerts

Rubakhin

et al. 2014

Analyst

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“…In these studies, Escherichia coli and Saccharomyces cerevisiae have frequently been employed as model microbes [5,6], because these species are biologically well characterized, and it is relatively easy to manipulate their genes. Mechanistic studies have revealed that HP treatment damages cellular organelles, membrane structures, and membrane proteins [6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Clarification of the recovery mechanism ofEscherichia coliafter hydrostatic pressure treatment

Ohshima

Nomura

Iwahashi

2013

High Pressure Research

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High hydrostatic pressure (HP) technology has gained more attention as a non-thermal food pasteurization technology. Recently, a limitation of the HP technology was reported by Koseki and Yamamoto [Recovery of Escherichia coli ATCC 25922 in phosphate buffered saline after treatment with high hydrostatic pressure. Int. J. Food Microbiol. 2006;110:108-111], who completely recovered Escherichia coli species after HP treatment. We investigated the recovery mechanism of E. coli after HP treatment. The cells were treated with 200-300 MPa at 0-25 • C for 24 h. The HP-treated E. coli was recovered in phosphate-buffered saline (PBS) during 120 h of incubation at 25 • C, confirming the results reported by them. However, E. coli did not grow in PBS but grew with inactivated cells in PBS. In addition, the results of our "population size experiments" demonstrated that the recovery of E. coli cells depended on both the degree of pressure and the population size. These results suggest that some portion of cells recovered from the damage and then grew by using inactivated cells.

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“…We assessed the metabolites of the cells restoring the damages after high-pressure treatment using CE/MS methods. We focused on amino acids, since these metabolites often contribute to the homeostasis of cells and can be easily separated from cells (Tanaka et al 2008). The metabolomics analysis suggested the significant accumulation of glycine, valine, isoleucine, leucine, asparagine, aspartic acid, and tyrosine in the cells (Tanaka et al 2010).…”

Section: Proteasome and Its Function Could Be The Crucial Target Of Dmentioning

confidence: 99%

“…Metabolomics is an emerging field in analytical biochemistry, and the development of this field for comprehensive and quantitative analysis of organic acids, carbohydrates, and nucleotides is a necessity in the field of functional genomics (Tanaka et al 2008). We assessed the metabolites of the cells restoring the damages after high-pressure treatment using CE/MS methods.…”

Section: Proteasome and Its Function Could Be The Crucial Target Of Dmentioning

confidence: 99%

Pressure-Dependent Gene Activation in Yeast Cells

Iwahashi

2015

Subcellular Biochemistry

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Hydrostatic pressure is one of the physical factors affecting cellular physiology. Hydrostatic pressure of a few tens MPa decreases the growth rate and a few hundred MPa decreases the cellular viability. To find clues to understand how such pressure effects on living cells relating to damages on protein molecules, we employed yeast DNA microarrays and analyzed genome-wide gene-expression levels in yeast cells which have been exposed to different levels of hydrostatic pressure. These include the cells temporarily adapted to a high pressure (from 0.1 to 30 MPa) and to a low pressure (from 30 to 0.1 MPa). These conditions cause yeast cells decreases of growth rate. We also analyzed gene expression profiles from the cells recovering after the sublethal pressure treatment at 180 MPa at 4 °C for 0 min and at 40 MPa at 4 °C for 16 h. These conditions cause yeast cells decreases of cellular viability. The activated genes by the temporary adaptations to both of the high pressure and the low pressure suggest that proteins related to membrane biosynthesis and cell wall biosynthesis can be crucial targets of pressure-induced damages, whereas the activated genes under recovering conditions after exposure to the sublethal high pressure suggest that proteasome activity and proteins localized in endoplasmic reticulum can be the crucial targets or the essential factors to survive.

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Development of a capillary electrophoresis‐mass spectrometry method using polymer capillaries for metabolomic analysis of yeast

Cited by 29 publications

References 31 publications

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Clarification of the recovery mechanism ofEscherichia coliafter hydrostatic pressure treatment

Pressure-Dependent Gene Activation in Yeast Cells

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