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A proteomics analysis to evaluate cytotoxicity in NRK-52E cells caused by unmodified Nano-Fe₃O₄.

Lin YR, Kuo CJ, Lin HY, Wu CJ, Liang SS - ScientificWorldJournal (2014)

Bottom Line: Through global proteomics analysis using dimethyl labeling techniques and liquid phase chromatography coupled with a tandem mass spectrometer (LC-MS/MS), we characterized 435 proteins including the programmed cell death related proteins, ras-related proteins, glutathione related proteins, and the chaperone proteins such as heat shock proteins, serpin H1, protein disulfide-isomerase A4, endoplasmin, and endoplasmic reticulum resident proteins.From the statistical data of identified proteins, we believed that NPs treatment causes cell death and promotes expression of ras-related proteins.In order to avoid apoptosis, NRK-52E cell lines induce a series of protective effects such as glutathione related proteins to reduce reactive oxygen species (ROS), and chaperone proteins to recycle damaged proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Fooyin University, 151 Jinxue Road, Kaohsiung 83102, Taiwan.

ABSTRACT
We synthesized unmodified Fe₃O₄ nanoparticles (NPs) with particles size from 10 nm to 100 nm. We cultured NRK-52E cell lines (rat, kidney) and treated with Fe₃O₄ NPs to investigate and evaluate the cytotoxicity of NPs for NRK-52E cells. Through global proteomics analysis using dimethyl labeling techniques and liquid phase chromatography coupled with a tandem mass spectrometer (LC-MS/MS), we characterized 435 proteins including the programmed cell death related proteins, ras-related proteins, glutathione related proteins, and the chaperone proteins such as heat shock proteins, serpin H1, protein disulfide-isomerase A4, endoplasmin, and endoplasmic reticulum resident proteins. From the statistical data of identified proteins, we believed that NPs treatment causes cell death and promotes expression of ras-related proteins. In order to avoid apoptosis, NRK-52E cell lines induce a series of protective effects such as glutathione related proteins to reduce reactive oxygen species (ROS), and chaperone proteins to recycle damaged proteins. We suggested that, in the indigenous cellular environment, Fe₃O₄ NPs treatment induced an antagonistic effect for cell lines to go to which avoids apoptosis.

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Related in: MedlinePlus

(a) Extracted ions of m/z 1165.92 (H-labeled) and m/z 1165.92 (D-labeled) showing signals of peak area (AA) and peak height (AH). (b) MS spectrum of GVDNTFADELVELSTALEHQEYITFLEDLK which are labeled formaldehyde-H2 and formaldehyde-D2. These two labeled peptides are shown in a coeluted isotopic pattern.
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fig4: (a) Extracted ions of m/z 1165.92 (H-labeled) and m/z 1165.92 (D-labeled) showing signals of peak area (AA) and peak height (AH). (b) MS spectrum of GVDNTFADELVELSTALEHQEYITFLEDLK which are labeled formaldehyde-H2 and formaldehyde-D2. These two labeled peptides are shown in a coeluted isotopic pattern.

Mentions: From the statistical results we identified 435 proteins, of which 311 proteins have a D/H ratio and 124 proteins which we were unable to specific values. For example, complement component 1 Q subcomponent-binding protein (C1qBP) was identified with an arithmetic average D/H ratio of 4.23. Figure 3 showed the peptide belonging to C1qBP having sequences GVDNTFADELVELSTALEHQEYITFLEDLK which was fragmentized by collision induced dissociation (CID), and its MS/MS spectrum was illustrated by y-ions and b-ions. From a quantitative standpoint in support of the Mascot Distiller in bioinformatics software, the result of protein list showed that the D/H ratio belonging to GVDNTFADELVELSTALEHQEYITFLEDLK peptide was 4.23. Simultaneously the raw data from LC-MS/MS that we extracted from the labeled peptide of C1qBP by m/z 1165.92 (3+, H-labeled) and 1168.60 (3+, D-labeled) presented signals of peak area and peak height (Figure 4(a)) and showed the isotope pattern of H-labeled peptide and D-labeled peptide (Figure 4(b)) which demonstrated the changes in different labeling. The statistical results showed that peak area was 4.42-fold in D/H ratio and peak height was 4.22-fold; the statistical data was calculated and shown in Table 1.


A proteomics analysis to evaluate cytotoxicity in NRK-52E cells caused by unmodified Nano-Fe₃O₄.

Lin YR, Kuo CJ, Lin HY, Wu CJ, Liang SS - ScientificWorldJournal (2014)

(a) Extracted ions of m/z 1165.92 (H-labeled) and m/z 1165.92 (D-labeled) showing signals of peak area (AA) and peak height (AH). (b) MS spectrum of GVDNTFADELVELSTALEHQEYITFLEDLK which are labeled formaldehyde-H2 and formaldehyde-D2. These two labeled peptides are shown in a coeluted isotopic pattern.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4150542&req=5

fig4: (a) Extracted ions of m/z 1165.92 (H-labeled) and m/z 1165.92 (D-labeled) showing signals of peak area (AA) and peak height (AH). (b) MS spectrum of GVDNTFADELVELSTALEHQEYITFLEDLK which are labeled formaldehyde-H2 and formaldehyde-D2. These two labeled peptides are shown in a coeluted isotopic pattern.
Mentions: From the statistical results we identified 435 proteins, of which 311 proteins have a D/H ratio and 124 proteins which we were unable to specific values. For example, complement component 1 Q subcomponent-binding protein (C1qBP) was identified with an arithmetic average D/H ratio of 4.23. Figure 3 showed the peptide belonging to C1qBP having sequences GVDNTFADELVELSTALEHQEYITFLEDLK which was fragmentized by collision induced dissociation (CID), and its MS/MS spectrum was illustrated by y-ions and b-ions. From a quantitative standpoint in support of the Mascot Distiller in bioinformatics software, the result of protein list showed that the D/H ratio belonging to GVDNTFADELVELSTALEHQEYITFLEDLK peptide was 4.23. Simultaneously the raw data from LC-MS/MS that we extracted from the labeled peptide of C1qBP by m/z 1165.92 (3+, H-labeled) and 1168.60 (3+, D-labeled) presented signals of peak area and peak height (Figure 4(a)) and showed the isotope pattern of H-labeled peptide and D-labeled peptide (Figure 4(b)) which demonstrated the changes in different labeling. The statistical results showed that peak area was 4.42-fold in D/H ratio and peak height was 4.22-fold; the statistical data was calculated and shown in Table 1.

Bottom Line: Through global proteomics analysis using dimethyl labeling techniques and liquid phase chromatography coupled with a tandem mass spectrometer (LC-MS/MS), we characterized 435 proteins including the programmed cell death related proteins, ras-related proteins, glutathione related proteins, and the chaperone proteins such as heat shock proteins, serpin H1, protein disulfide-isomerase A4, endoplasmin, and endoplasmic reticulum resident proteins.From the statistical data of identified proteins, we believed that NPs treatment causes cell death and promotes expression of ras-related proteins.In order to avoid apoptosis, NRK-52E cell lines induce a series of protective effects such as glutathione related proteins to reduce reactive oxygen species (ROS), and chaperone proteins to recycle damaged proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Fooyin University, 151 Jinxue Road, Kaohsiung 83102, Taiwan.

ABSTRACT
We synthesized unmodified Fe₃O₄ nanoparticles (NPs) with particles size from 10 nm to 100 nm. We cultured NRK-52E cell lines (rat, kidney) and treated with Fe₃O₄ NPs to investigate and evaluate the cytotoxicity of NPs for NRK-52E cells. Through global proteomics analysis using dimethyl labeling techniques and liquid phase chromatography coupled with a tandem mass spectrometer (LC-MS/MS), we characterized 435 proteins including the programmed cell death related proteins, ras-related proteins, glutathione related proteins, and the chaperone proteins such as heat shock proteins, serpin H1, protein disulfide-isomerase A4, endoplasmin, and endoplasmic reticulum resident proteins. From the statistical data of identified proteins, we believed that NPs treatment causes cell death and promotes expression of ras-related proteins. In order to avoid apoptosis, NRK-52E cell lines induce a series of protective effects such as glutathione related proteins to reduce reactive oxygen species (ROS), and chaperone proteins to recycle damaged proteins. We suggested that, in the indigenous cellular environment, Fe₃O₄ NPs treatment induced an antagonistic effect for cell lines to go to which avoids apoptosis.

Show MeSH
Related in: MedlinePlus