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S-Nitrosylation in Organs of Mice Exposed to Low or High Doses of γ-Rays: The Modulating Effect of Iodine Contrast Agent at a Low Radiation Dose.

Nicolas F, Wu C, Bukhari S, de Toledo SM, Li H, Shibata M, Azzam EI - Proteomes (2015)

Bottom Line: The covalent addition of nitric oxide (NO(•)) onto cysteine thiols, or S-nitrosylation, modulates the activity of key signaling proteins.To gain insight into the biochemical changes induced by low-dose ionizing radiation, we determined global S-nitrosylation by the "biotin switch" assay coupled with mass spectrometry analyses in organs of C57BL/6J mice exposed to acute 0.1 Gy of (137)Cs γ-rays.The major results were as follows: (i) iopamidol alone had significant effects on S-nitrosylation in brain, lung and liver; (ii) relative to the control, exposure to 0.1 Gy without iopamidol resulted in statistically-significant SNO changes in proteins that differ in molecular weight in liver, lung, brain and blood plasma; (iii) iopamidol enhanced the decrease in S-nitrosylation induced by 0.1 Gy in brain; (iv) whereas a decrease in S-nitrosylation occurred at 0.1 Gy for proteins of ~50 kDa in brain and for proteins of ~37 kDa in liver, an increase was detected at 4 Gy in both organs; (v) mass spectrometry analyses of nitrosylated proteins in brain revealed differential modulation of SNO proteins (e.g., sodium/potassium-transporting ATPase subunit beta-1; beta tubulins; ADP-ribosylation factor 5) by low- and high-dose irradiation; and (vi) ingenuity pathway analysis identified major signaling networks to be modulated, in particular the neuronal nitric oxide synthase signaling pathway was differentially modulated by low- and high-dose γ-irradiation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Health Informatics, Rutgers School of Health Related Professions, Newark, NJ 07107, USA; nicolafa@shrp.rutgers.edu.

ABSTRACT

The covalent addition of nitric oxide (NO(•)) onto cysteine thiols, or S-nitrosylation, modulates the activity of key signaling proteins. The dysregulation of normal S-nitrosylation contributes to degenerative conditions and to cancer. To gain insight into the biochemical changes induced by low-dose ionizing radiation, we determined global S-nitrosylation by the "biotin switch" assay coupled with mass spectrometry analyses in organs of C57BL/6J mice exposed to acute 0.1 Gy of (137)Cs γ-rays. The dose of radiation was delivered to the whole body in the presence or absence of iopamidol, an iodinated contrast agent used during radiological examinations. To investigate whether similar or distinct nitrosylation patterns are induced following high-dose irradiation, mice were exposed in parallel to acute 4 Gy of (137)Cs γ rays. Analysis of modulated S-nitrosothiols (SNO-proteins) in freshly-harvested organs of animals sacrificed 13 days after irradiation revealed radiation dose- and contrast agent-dependent changes. The major results were as follows: (i) iopamidol alone had significant effects on S-nitrosylation in brain, lung and liver; (ii) relative to the control, exposure to 0.1 Gy without iopamidol resulted in statistically-significant SNO changes in proteins that differ in molecular weight in liver, lung, brain and blood plasma; (iii) iopamidol enhanced the decrease in S-nitrosylation induced by 0.1 Gy in brain; (iv) whereas a decrease in S-nitrosylation occurred at 0.1 Gy for proteins of ~50 kDa in brain and for proteins of ~37 kDa in liver, an increase was detected at 4 Gy in both organs; (v) mass spectrometry analyses of nitrosylated proteins in brain revealed differential modulation of SNO proteins (e.g., sodium/potassium-transporting ATPase subunit beta-1; beta tubulins; ADP-ribosylation factor 5) by low- and high-dose irradiation; and (vi) ingenuity pathway analysis identified major signaling networks to be modulated, in particular the neuronal nitric oxide synthase signaling pathway was differentially modulated by low- and high-dose γ-irradiation.

No MeSH data available.


Related in: MedlinePlus

Differential effect of γ-ray dose on S-nitrosylation level. A comparison of spectral counts of SNO proteins (proteins with spectral counts above 80) showed differential modulation relative to the control of β tubulins in brain of mice exposed to 4 Gy, while no such effect was observed at 0.1 Gy. S-nitrosylation of ADP-ribosylation factor 5 (ARF5 shown in green) increased at 4 Gy.
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Figure 4: Differential effect of γ-ray dose on S-nitrosylation level. A comparison of spectral counts of SNO proteins (proteins with spectral counts above 80) showed differential modulation relative to the control of β tubulins in brain of mice exposed to 4 Gy, while no such effect was observed at 0.1 Gy. S-nitrosylation of ADP-ribosylation factor 5 (ARF5 shown in green) increased at 4 Gy.

Mentions: The majority of the detected proteins had spectral counts of less than 10, and there were only small deviations among the three different conditions. Only a few proteins exhibited a spectral count above 80 regardless of the experimental conditions (Figure 4). Among these six proteins, tubulin betas (TBB4 and TBB2A) showed a slight decrease in spectral counts at 0.1 Gy compared to 0 Gy, while the magnitude of decrease was more enhanced at 4 Gy (Figure 4, line graph or, alternatively, the bar graph in Supplementary Figure S2). Compared to tubulin beta, no appreciable changes were observed for tubulin alphas (TBA1A, TBA1B, TBA4A) at both low and high doses compared to 0 Gy. One protein that showed almost no difference in spectral counts at 0.1 Gy, but a slight increase at 4 Gy, is ADP-ribosylation factor 5 (ARF5) (Figure 4). Another protein that exhibited a >80 spectral count is AT1B1 (sodium/potassium-transporting ATPase subunit β-1). Compared to the other six proteins, AT1B1 showed spectral counts of 25 and 20 at 0 Gy and 4 Gy, respectively, while the count rose to 120 at 0.1 Gy.


S-Nitrosylation in Organs of Mice Exposed to Low or High Doses of γ-Rays: The Modulating Effect of Iodine Contrast Agent at a Low Radiation Dose.

Nicolas F, Wu C, Bukhari S, de Toledo SM, Li H, Shibata M, Azzam EI - Proteomes (2015)

Differential effect of γ-ray dose on S-nitrosylation level. A comparison of spectral counts of SNO proteins (proteins with spectral counts above 80) showed differential modulation relative to the control of β tubulins in brain of mice exposed to 4 Gy, while no such effect was observed at 0.1 Gy. S-nitrosylation of ADP-ribosylation factor 5 (ARF5 shown in green) increased at 4 Gy.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4548934&req=5

Figure 4: Differential effect of γ-ray dose on S-nitrosylation level. A comparison of spectral counts of SNO proteins (proteins with spectral counts above 80) showed differential modulation relative to the control of β tubulins in brain of mice exposed to 4 Gy, while no such effect was observed at 0.1 Gy. S-nitrosylation of ADP-ribosylation factor 5 (ARF5 shown in green) increased at 4 Gy.
Mentions: The majority of the detected proteins had spectral counts of less than 10, and there were only small deviations among the three different conditions. Only a few proteins exhibited a spectral count above 80 regardless of the experimental conditions (Figure 4). Among these six proteins, tubulin betas (TBB4 and TBB2A) showed a slight decrease in spectral counts at 0.1 Gy compared to 0 Gy, while the magnitude of decrease was more enhanced at 4 Gy (Figure 4, line graph or, alternatively, the bar graph in Supplementary Figure S2). Compared to tubulin beta, no appreciable changes were observed for tubulin alphas (TBA1A, TBA1B, TBA4A) at both low and high doses compared to 0 Gy. One protein that showed almost no difference in spectral counts at 0.1 Gy, but a slight increase at 4 Gy, is ADP-ribosylation factor 5 (ARF5) (Figure 4). Another protein that exhibited a >80 spectral count is AT1B1 (sodium/potassium-transporting ATPase subunit β-1). Compared to the other six proteins, AT1B1 showed spectral counts of 25 and 20 at 0 Gy and 4 Gy, respectively, while the count rose to 120 at 0.1 Gy.

Bottom Line: The covalent addition of nitric oxide (NO(•)) onto cysteine thiols, or S-nitrosylation, modulates the activity of key signaling proteins.To gain insight into the biochemical changes induced by low-dose ionizing radiation, we determined global S-nitrosylation by the "biotin switch" assay coupled with mass spectrometry analyses in organs of C57BL/6J mice exposed to acute 0.1 Gy of (137)Cs γ-rays.The major results were as follows: (i) iopamidol alone had significant effects on S-nitrosylation in brain, lung and liver; (ii) relative to the control, exposure to 0.1 Gy without iopamidol resulted in statistically-significant SNO changes in proteins that differ in molecular weight in liver, lung, brain and blood plasma; (iii) iopamidol enhanced the decrease in S-nitrosylation induced by 0.1 Gy in brain; (iv) whereas a decrease in S-nitrosylation occurred at 0.1 Gy for proteins of ~50 kDa in brain and for proteins of ~37 kDa in liver, an increase was detected at 4 Gy in both organs; (v) mass spectrometry analyses of nitrosylated proteins in brain revealed differential modulation of SNO proteins (e.g., sodium/potassium-transporting ATPase subunit beta-1; beta tubulins; ADP-ribosylation factor 5) by low- and high-dose irradiation; and (vi) ingenuity pathway analysis identified major signaling networks to be modulated, in particular the neuronal nitric oxide synthase signaling pathway was differentially modulated by low- and high-dose γ-irradiation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Health Informatics, Rutgers School of Health Related Professions, Newark, NJ 07107, USA; nicolafa@shrp.rutgers.edu.

ABSTRACT

The covalent addition of nitric oxide (NO(•)) onto cysteine thiols, or S-nitrosylation, modulates the activity of key signaling proteins. The dysregulation of normal S-nitrosylation contributes to degenerative conditions and to cancer. To gain insight into the biochemical changes induced by low-dose ionizing radiation, we determined global S-nitrosylation by the "biotin switch" assay coupled with mass spectrometry analyses in organs of C57BL/6J mice exposed to acute 0.1 Gy of (137)Cs γ-rays. The dose of radiation was delivered to the whole body in the presence or absence of iopamidol, an iodinated contrast agent used during radiological examinations. To investigate whether similar or distinct nitrosylation patterns are induced following high-dose irradiation, mice were exposed in parallel to acute 4 Gy of (137)Cs γ rays. Analysis of modulated S-nitrosothiols (SNO-proteins) in freshly-harvested organs of animals sacrificed 13 days after irradiation revealed radiation dose- and contrast agent-dependent changes. The major results were as follows: (i) iopamidol alone had significant effects on S-nitrosylation in brain, lung and liver; (ii) relative to the control, exposure to 0.1 Gy without iopamidol resulted in statistically-significant SNO changes in proteins that differ in molecular weight in liver, lung, brain and blood plasma; (iii) iopamidol enhanced the decrease in S-nitrosylation induced by 0.1 Gy in brain; (iv) whereas a decrease in S-nitrosylation occurred at 0.1 Gy for proteins of ~50 kDa in brain and for proteins of ~37 kDa in liver, an increase was detected at 4 Gy in both organs; (v) mass spectrometry analyses of nitrosylated proteins in brain revealed differential modulation of SNO proteins (e.g., sodium/potassium-transporting ATPase subunit beta-1; beta tubulins; ADP-ribosylation factor 5) by low- and high-dose irradiation; and (vi) ingenuity pathway analysis identified major signaling networks to be modulated, in particular the neuronal nitric oxide synthase signaling pathway was differentially modulated by low- and high-dose γ-irradiation.

No MeSH data available.


Related in: MedlinePlus