Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase: Implications for enzymatic activity and antioxidant action

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase : Implications for enzymatic activity and antioxidant action. / Tiwari, Manish Kumar; Hägglund, Per Mårten; Møller, Ian Max; Davies, Michael Jonathan; Bjerrum, Morten Jannik.

In: Redox Biology, Vol. 26, 101262, 2019.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Tiwari, MK, Hägglund, PM, Møller, IM, Davies, MJ & Bjerrum, MJ 2019, 'Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase: Implications for enzymatic activity and antioxidant action', Redox Biology, vol. 26, 101262. https://doi.org/10.1016/j.redox.2019.101262

APA

Tiwari, M. K., Hägglund, P. M., Møller, I. M., Davies, M. J., & Bjerrum, M. J. (2019). Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase: Implications for enzymatic activity and antioxidant action. Redox Biology, 26, [101262]. https://doi.org/10.1016/j.redox.2019.101262

Vancouver

Tiwari MK, Hägglund PM, Møller IM, Davies MJ, Bjerrum MJ. Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase: Implications for enzymatic activity and antioxidant action. Redox Biology. 2019;26. 101262. https://doi.org/10.1016/j.redox.2019.101262

Author

Tiwari, Manish Kumar ; Hägglund, Per Mårten ; Møller, Ian Max ; Davies, Michael Jonathan ; Bjerrum, Morten Jannik. / Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase : Implications for enzymatic activity and antioxidant action. In: Redox Biology. 2019 ; Vol. 26.

Bibtex

@article{6269c9b0049a4db5b04f4f3fccf5cd45,
title = "Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase: Implications for enzymatic activity and antioxidant action",
abstract = "Copper ion-catalyzed oxidation of yeast SOD1 (ySOD1) was examined to determine early oxidative modifications, including oxidation of a crucial disulfide bond, and the structural and functional repercussions of these events. The study used distinct oxidative conditions: Cu2+/H2O2, Cu2+/H2O2/AscH(-) and Cu2+/H2O2/glucose. Capillary electrophoresis experiments and quantification of protein carbonyls indicate that ySOD1 is highly susceptible to oxidative modification and that changes can be detected within 0.1 min of the initiation of the reaction. Oxidation-induced structural perturbations, characterized by circular dichroism, revealed the formation of partially-unfolded ySOD1 species in a dose-dependent manner. Consistent with these structural changes, pyrogallol assay indicates a partial loss of enzymatic activity. ESI-MS analyses showed seven distinct oxidized ySOD1 species under mild oxidation within 0.1 min. LC/MS analysis after proteolytic digestion demonstrated that the copper-coordinating active site histidine residues, His47 and His49, were converted into 2-oxo-histidine. Furthermore, the Cu and Zn bridging residue, His64 is converted into aspartate/asparagine. Importantly, the disulfide-bond Cys58-Cys147 which is critical for the structural and functional integrity of ySOD1 was detected as being oxidized at Cys147. We propose, based on LC/MS analyses, that disulfide-bond oxidation occurs without disulfide bond cleavage. Modifications were also detected at Met85 and five surface-exposed Lys residues. Based on these data we propose that the Cys58-Cys147 bond may act as a sacrificial target for oxidants and protect ySOD1 from oxidative inactivation arising from exposure to Cu2+/H2O2 and auto-inactivation during extended enzymatic turnover.",
author = "Tiwari, {Manish Kumar} and H{\"a}gglund, {Per M{\aa}rten} and M{\o}ller, {Ian Max} and Davies, {Michael Jonathan} and Bjerrum, {Morten Jannik}",
year = "2019",
doi = "10.1016/j.redox.2019.101262",
language = "English",
volume = "26",
journal = "Redox Biology",
issn = "2213-2317",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Copper ion / H2O2 oxidation of Cu/Zn-Superoxide dismutase

T2 - Implications for enzymatic activity and antioxidant action

AU - Tiwari, Manish Kumar

AU - Hägglund, Per Mårten

AU - Møller, Ian Max

AU - Davies, Michael Jonathan

AU - Bjerrum, Morten Jannik

PY - 2019

Y1 - 2019

N2 - Copper ion-catalyzed oxidation of yeast SOD1 (ySOD1) was examined to determine early oxidative modifications, including oxidation of a crucial disulfide bond, and the structural and functional repercussions of these events. The study used distinct oxidative conditions: Cu2+/H2O2, Cu2+/H2O2/AscH(-) and Cu2+/H2O2/glucose. Capillary electrophoresis experiments and quantification of protein carbonyls indicate that ySOD1 is highly susceptible to oxidative modification and that changes can be detected within 0.1 min of the initiation of the reaction. Oxidation-induced structural perturbations, characterized by circular dichroism, revealed the formation of partially-unfolded ySOD1 species in a dose-dependent manner. Consistent with these structural changes, pyrogallol assay indicates a partial loss of enzymatic activity. ESI-MS analyses showed seven distinct oxidized ySOD1 species under mild oxidation within 0.1 min. LC/MS analysis after proteolytic digestion demonstrated that the copper-coordinating active site histidine residues, His47 and His49, were converted into 2-oxo-histidine. Furthermore, the Cu and Zn bridging residue, His64 is converted into aspartate/asparagine. Importantly, the disulfide-bond Cys58-Cys147 which is critical for the structural and functional integrity of ySOD1 was detected as being oxidized at Cys147. We propose, based on LC/MS analyses, that disulfide-bond oxidation occurs without disulfide bond cleavage. Modifications were also detected at Met85 and five surface-exposed Lys residues. Based on these data we propose that the Cys58-Cys147 bond may act as a sacrificial target for oxidants and protect ySOD1 from oxidative inactivation arising from exposure to Cu2+/H2O2 and auto-inactivation during extended enzymatic turnover.

AB - Copper ion-catalyzed oxidation of yeast SOD1 (ySOD1) was examined to determine early oxidative modifications, including oxidation of a crucial disulfide bond, and the structural and functional repercussions of these events. The study used distinct oxidative conditions: Cu2+/H2O2, Cu2+/H2O2/AscH(-) and Cu2+/H2O2/glucose. Capillary electrophoresis experiments and quantification of protein carbonyls indicate that ySOD1 is highly susceptible to oxidative modification and that changes can be detected within 0.1 min of the initiation of the reaction. Oxidation-induced structural perturbations, characterized by circular dichroism, revealed the formation of partially-unfolded ySOD1 species in a dose-dependent manner. Consistent with these structural changes, pyrogallol assay indicates a partial loss of enzymatic activity. ESI-MS analyses showed seven distinct oxidized ySOD1 species under mild oxidation within 0.1 min. LC/MS analysis after proteolytic digestion demonstrated that the copper-coordinating active site histidine residues, His47 and His49, were converted into 2-oxo-histidine. Furthermore, the Cu and Zn bridging residue, His64 is converted into aspartate/asparagine. Importantly, the disulfide-bond Cys58-Cys147 which is critical for the structural and functional integrity of ySOD1 was detected as being oxidized at Cys147. We propose, based on LC/MS analyses, that disulfide-bond oxidation occurs without disulfide bond cleavage. Modifications were also detected at Met85 and five surface-exposed Lys residues. Based on these data we propose that the Cys58-Cys147 bond may act as a sacrificial target for oxidants and protect ySOD1 from oxidative inactivation arising from exposure to Cu2+/H2O2 and auto-inactivation during extended enzymatic turnover.

U2 - 10.1016/j.redox.2019.101262

DO - 10.1016/j.redox.2019.101262

M3 - Journal article

VL - 26

JO - Redox Biology

JF - Redox Biology

SN - 2213-2317

M1 - 101262

ER -

ID: 224548456