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Fasting, but Not Aging, Dramatically Alters the Redox Status of Cysteine Residues on Proteins in Drosophila melanogaster.

Menger KE, James AM, Cochemé HM, Harbour ME, Chouchani ET, Ding S, Fearnley IM, Partridge L, Murphy MP - Cell Rep (2015)

Bottom Line: Surprisingly, these cysteine residues did not become more oxidized with age.In contrast, 24 hr of fasting dramatically oxidized cysteine residues that were reduced under fed conditions while also reducing cysteine residues that were initially oxidized.We conclude that fasting, but not aging, dramatically alters cysteine-residue redox status in D. melanogaster.

View Article: PubMed Central - PubMed

Affiliation: MRC Mitochondrial Biology Unit, Cambridge CB2 0XY, UK; Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

No MeSH data available.


Related in: MedlinePlus

Effect of Exogenous Oxidants on Protein Cysteine-Residue Redox State and Fly Survival(A) Survival of young (7 days) control and catalase-overexpressing flies after exposure to H2O2. Arrow indicates when cohorts are collected (24-hr treatment).(B) Distribution of cysteine peptides plotted against redox states of the cysteine residues for control flies after exposure to H2O2. Data show the mean of five biological samples where each cysteine residue identified is sorted into corresponding 5% quantiles, and the resulting distributions are averaged (mean ± SEM). Dashed line indicates the untreated control (cf. Figure 2B).(C) Oxidation state of cysteine residues present in at least three biological replicates exposed to H2O2 plotted against the same cysteine residues present in at least three biological replicates of controls. Dotted line slope = 1, whereas the continuous line is the best fit to the data. Red symbols (n = 12) indicate cysteine residues significantly different following a non-paired, two-tailed Student’s t test (p < 0.05). Total unique peptides = 452.(D) Survival of young (7 days) control and catalase overexpressing flies after exposure to PQ. Arrow indicates where cohorts are sampled (24-hr treatment).(E) Distribution of cysteine-containing peptides plotted against redox states of the cysteine residues for control flies after exposure to PQ. Means are across five biological replicates of the relative number of cysteine residues within each 5% quantile. The dashed line is the untreated control cohort (cf. Figure 2B).(F) Oxidation state of cysteine residues in control flies exposed to PQ plotted against untreated flies. Dotted line slope = 1, while the continuous line is the line of best fit. Each symbol represents a cysteine residue identified in at least three biological replicates of the untreated as well as the PQ-treated cohort. Red symbols identify cysteine residues (n = 68; p < 0.05; non-paired, two-tailed Student’s t test with low-stringency significance). The blue symbols (n = 33) indicate a high-stringency significance (Benjamini-Hochberg test). Total unique peptides = 452.See also Figure S4.
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fig4: Effect of Exogenous Oxidants on Protein Cysteine-Residue Redox State and Fly Survival(A) Survival of young (7 days) control and catalase-overexpressing flies after exposure to H2O2. Arrow indicates when cohorts are collected (24-hr treatment).(B) Distribution of cysteine peptides plotted against redox states of the cysteine residues for control flies after exposure to H2O2. Data show the mean of five biological samples where each cysteine residue identified is sorted into corresponding 5% quantiles, and the resulting distributions are averaged (mean ± SEM). Dashed line indicates the untreated control (cf. Figure 2B).(C) Oxidation state of cysteine residues present in at least three biological replicates exposed to H2O2 plotted against the same cysteine residues present in at least three biological replicates of controls. Dotted line slope = 1, whereas the continuous line is the best fit to the data. Red symbols (n = 12) indicate cysteine residues significantly different following a non-paired, two-tailed Student’s t test (p < 0.05). Total unique peptides = 452.(D) Survival of young (7 days) control and catalase overexpressing flies after exposure to PQ. Arrow indicates where cohorts are sampled (24-hr treatment).(E) Distribution of cysteine-containing peptides plotted against redox states of the cysteine residues for control flies after exposure to PQ. Means are across five biological replicates of the relative number of cysteine residues within each 5% quantile. The dashed line is the untreated control cohort (cf. Figure 2B).(F) Oxidation state of cysteine residues in control flies exposed to PQ plotted against untreated flies. Dotted line slope = 1, while the continuous line is the line of best fit. Each symbol represents a cysteine residue identified in at least three biological replicates of the untreated as well as the PQ-treated cohort. Red symbols identify cysteine residues (n = 68; p < 0.05; non-paired, two-tailed Student’s t test with low-stringency significance). The blue symbols (n = 33) indicate a high-stringency significance (Benjamini-Hochberg test). Total unique peptides = 452.See also Figure S4.

Mentions: Given that a lack of cysteine-residue oxidation with age was surprising, we investigated whether cysteine-residue oxidation responded to H2O2 in vivo. Dietary H2O2 dramatically decreased survival of control flies, and overexpressing catalase conferred resistance (Figure 4A). Next, we analyzed control untreated and H2O2-treated flies by OxICAT, and we found there was a marginal oxidation of the cysteine residues (26.3%; Figure 4B) in comparison to untreated control flies (22%; Figure 2B). The redox state of the individual cysteine residues following H2O2 treatment was then plotted against those in untreated control flies (Figure 4C). If H2O2 treatment did not affect cysteine-residue redox state, then the points would lie on the dotted line, and H2O2 shifted very few peptides above this line (Figure 4C). Overexpressing catalase had little effect on cysteine residue oxidation by H2O2 (Figures S4A and S4B). Thus, surprisingly, H2O2 levels that dramatically decrease survival did so without oxidizing cysteine residues (Figure 4C).


Fasting, but Not Aging, Dramatically Alters the Redox Status of Cysteine Residues on Proteins in Drosophila melanogaster.

Menger KE, James AM, Cochemé HM, Harbour ME, Chouchani ET, Ding S, Fearnley IM, Partridge L, Murphy MP - Cell Rep (2015)

Effect of Exogenous Oxidants on Protein Cysteine-Residue Redox State and Fly Survival(A) Survival of young (7 days) control and catalase-overexpressing flies after exposure to H2O2. Arrow indicates when cohorts are collected (24-hr treatment).(B) Distribution of cysteine peptides plotted against redox states of the cysteine residues for control flies after exposure to H2O2. Data show the mean of five biological samples where each cysteine residue identified is sorted into corresponding 5% quantiles, and the resulting distributions are averaged (mean ± SEM). Dashed line indicates the untreated control (cf. Figure 2B).(C) Oxidation state of cysteine residues present in at least three biological replicates exposed to H2O2 plotted against the same cysteine residues present in at least three biological replicates of controls. Dotted line slope = 1, whereas the continuous line is the best fit to the data. Red symbols (n = 12) indicate cysteine residues significantly different following a non-paired, two-tailed Student’s t test (p < 0.05). Total unique peptides = 452.(D) Survival of young (7 days) control and catalase overexpressing flies after exposure to PQ. Arrow indicates where cohorts are sampled (24-hr treatment).(E) Distribution of cysteine-containing peptides plotted against redox states of the cysteine residues for control flies after exposure to PQ. Means are across five biological replicates of the relative number of cysteine residues within each 5% quantile. The dashed line is the untreated control cohort (cf. Figure 2B).(F) Oxidation state of cysteine residues in control flies exposed to PQ plotted against untreated flies. Dotted line slope = 1, while the continuous line is the line of best fit. Each symbol represents a cysteine residue identified in at least three biological replicates of the untreated as well as the PQ-treated cohort. Red symbols identify cysteine residues (n = 68; p < 0.05; non-paired, two-tailed Student’s t test with low-stringency significance). The blue symbols (n = 33) indicate a high-stringency significance (Benjamini-Hochberg test). Total unique peptides = 452.See also Figure S4.
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fig4: Effect of Exogenous Oxidants on Protein Cysteine-Residue Redox State and Fly Survival(A) Survival of young (7 days) control and catalase-overexpressing flies after exposure to H2O2. Arrow indicates when cohorts are collected (24-hr treatment).(B) Distribution of cysteine peptides plotted against redox states of the cysteine residues for control flies after exposure to H2O2. Data show the mean of five biological samples where each cysteine residue identified is sorted into corresponding 5% quantiles, and the resulting distributions are averaged (mean ± SEM). Dashed line indicates the untreated control (cf. Figure 2B).(C) Oxidation state of cysteine residues present in at least three biological replicates exposed to H2O2 plotted against the same cysteine residues present in at least three biological replicates of controls. Dotted line slope = 1, whereas the continuous line is the best fit to the data. Red symbols (n = 12) indicate cysteine residues significantly different following a non-paired, two-tailed Student’s t test (p < 0.05). Total unique peptides = 452.(D) Survival of young (7 days) control and catalase overexpressing flies after exposure to PQ. Arrow indicates where cohorts are sampled (24-hr treatment).(E) Distribution of cysteine-containing peptides plotted against redox states of the cysteine residues for control flies after exposure to PQ. Means are across five biological replicates of the relative number of cysteine residues within each 5% quantile. The dashed line is the untreated control cohort (cf. Figure 2B).(F) Oxidation state of cysteine residues in control flies exposed to PQ plotted against untreated flies. Dotted line slope = 1, while the continuous line is the line of best fit. Each symbol represents a cysteine residue identified in at least three biological replicates of the untreated as well as the PQ-treated cohort. Red symbols identify cysteine residues (n = 68; p < 0.05; non-paired, two-tailed Student’s t test with low-stringency significance). The blue symbols (n = 33) indicate a high-stringency significance (Benjamini-Hochberg test). Total unique peptides = 452.See also Figure S4.
Mentions: Given that a lack of cysteine-residue oxidation with age was surprising, we investigated whether cysteine-residue oxidation responded to H2O2 in vivo. Dietary H2O2 dramatically decreased survival of control flies, and overexpressing catalase conferred resistance (Figure 4A). Next, we analyzed control untreated and H2O2-treated flies by OxICAT, and we found there was a marginal oxidation of the cysteine residues (26.3%; Figure 4B) in comparison to untreated control flies (22%; Figure 2B). The redox state of the individual cysteine residues following H2O2 treatment was then plotted against those in untreated control flies (Figure 4C). If H2O2 treatment did not affect cysteine-residue redox state, then the points would lie on the dotted line, and H2O2 shifted very few peptides above this line (Figure 4C). Overexpressing catalase had little effect on cysteine residue oxidation by H2O2 (Figures S4A and S4B). Thus, surprisingly, H2O2 levels that dramatically decrease survival did so without oxidizing cysteine residues (Figure 4C).

Bottom Line: Surprisingly, these cysteine residues did not become more oxidized with age.In contrast, 24 hr of fasting dramatically oxidized cysteine residues that were reduced under fed conditions while also reducing cysteine residues that were initially oxidized.We conclude that fasting, but not aging, dramatically alters cysteine-residue redox status in D. melanogaster.

View Article: PubMed Central - PubMed

Affiliation: MRC Mitochondrial Biology Unit, Cambridge CB2 0XY, UK; Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

No MeSH data available.


Related in: MedlinePlus