Irreversible inactivation of glutathione peroxidase 1 and reversible inactivation of peroxiredoxin ii by H2O2 in red blood cells

Chun Seok Cho, Sukmook Lee, Geun Taek Lee, Hyun Ae Woo, Eui Ju Choi, Sue Goo Rhee

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Catalase, glutathione peroxidase1 (GPx1), and peroxiredoxin (Prx) II are the principal enzymes responsible for peroxide elimination in RBC. We have now evaluated the relative roles of these enzymes by studying inactivation of GPx1 and Prx II in human RBCs. Mass spectrometry revealed that treatment of GPx1 with H2O2 converts the selenocysteine residue at its active site to dehydroalanine (DHA). We developed a blot method for detection of DHA-containing proteins, with which we observed that the amount of DHA-containing GPx1 increases with increasing RBC density, which is correlated with increasing RBC age. Given that the conversion of selenocysteine to DHA is irreversible, the content of DHA-GPx1 in each RBC likely reflects total oxidative stress experienced by the cell during its lifetime. Prx II is inactivated by occasional hyperoxidation of its catalytic cysteine to cysteine sulfinic acid during catalysis. We believe that the activity of sulfiredoxin in RBCs is sufficient to counteract the hyperoxidation of Prx II that occurs in the presence of the basal level of H2O2 flux resulting from hemoglobin autoxidation. If the H2O2 flux is increased above the basal level, however, the sulfinic Prx II begins to accumulate. In the presence of an increased H2O2 flux, inhibition of catalase accelerated the accumulation of sulfinic Prx II, indicative of the protective role of catalase. Antioxid. Redox Signal. 12, 1235-1246.

Original languageEnglish
Pages (from-to)1235-1246
Number of pages12
JournalAntioxidants and Redox Signaling
Volume12
Issue number11
DOIs
Publication statusPublished - 2010 Jun 1

Fingerprint

Peroxiredoxins
Blood
Erythrocytes
Glutathione
Cells
Catalase
Selenocysteine
Fluxes
Oxidative stress
Peroxides
Enzymes
Catalysis
Oxidation-Reduction
Mass spectrometry
Cysteine
glutathione peroxidase GPX1
Mass Spectrometry
Catalytic Domain
Hemoglobins
Oxidative Stress

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Physiology
  • Clinical Biochemistry

Cite this

Irreversible inactivation of glutathione peroxidase 1 and reversible inactivation of peroxiredoxin ii by H2O2 in red blood cells. / Cho, Chun Seok; Lee, Sukmook; Lee, Geun Taek; Woo, Hyun Ae; Choi, Eui Ju; Rhee, Sue Goo.

In: Antioxidants and Redox Signaling, Vol. 12, No. 11, 01.06.2010, p. 1235-1246.

Research output: Contribution to journalArticle

Cho, Chun Seok ; Lee, Sukmook ; Lee, Geun Taek ; Woo, Hyun Ae ; Choi, Eui Ju ; Rhee, Sue Goo. / Irreversible inactivation of glutathione peroxidase 1 and reversible inactivation of peroxiredoxin ii by H2O2 in red blood cells. In: Antioxidants and Redox Signaling. 2010 ; Vol. 12, No. 11. pp. 1235-1246.
@article{639df81b587841cdb9a4cff96ea87c90,
title = "Irreversible inactivation of glutathione peroxidase 1 and reversible inactivation of peroxiredoxin ii by H2O2 in red blood cells",
abstract = "Catalase, glutathione peroxidase1 (GPx1), and peroxiredoxin (Prx) II are the principal enzymes responsible for peroxide elimination in RBC. We have now evaluated the relative roles of these enzymes by studying inactivation of GPx1 and Prx II in human RBCs. Mass spectrometry revealed that treatment of GPx1 with H2O2 converts the selenocysteine residue at its active site to dehydroalanine (DHA). We developed a blot method for detection of DHA-containing proteins, with which we observed that the amount of DHA-containing GPx1 increases with increasing RBC density, which is correlated with increasing RBC age. Given that the conversion of selenocysteine to DHA is irreversible, the content of DHA-GPx1 in each RBC likely reflects total oxidative stress experienced by the cell during its lifetime. Prx II is inactivated by occasional hyperoxidation of its catalytic cysteine to cysteine sulfinic acid during catalysis. We believe that the activity of sulfiredoxin in RBCs is sufficient to counteract the hyperoxidation of Prx II that occurs in the presence of the basal level of H2O2 flux resulting from hemoglobin autoxidation. If the H2O2 flux is increased above the basal level, however, the sulfinic Prx II begins to accumulate. In the presence of an increased H2O2 flux, inhibition of catalase accelerated the accumulation of sulfinic Prx II, indicative of the protective role of catalase. Antioxid. Redox Signal. 12, 1235-1246.",
author = "Cho, {Chun Seok} and Sukmook Lee and Lee, {Geun Taek} and Woo, {Hyun Ae} and Choi, {Eui Ju} and Rhee, {Sue Goo}",
year = "2010",
month = "6",
day = "1",
doi = "10.1089/ars.2009.2701",
language = "English",
volume = "12",
pages = "1235--1246",
journal = "Antioxidants and Redox Signaling",
issn = "1523-0864",
publisher = "Mary Ann Liebert Inc.",
number = "11",

}

TY - JOUR

T1 - Irreversible inactivation of glutathione peroxidase 1 and reversible inactivation of peroxiredoxin ii by H2O2 in red blood cells

AU - Cho, Chun Seok

AU - Lee, Sukmook

AU - Lee, Geun Taek

AU - Woo, Hyun Ae

AU - Choi, Eui Ju

AU - Rhee, Sue Goo

PY - 2010/6/1

Y1 - 2010/6/1

N2 - Catalase, glutathione peroxidase1 (GPx1), and peroxiredoxin (Prx) II are the principal enzymes responsible for peroxide elimination in RBC. We have now evaluated the relative roles of these enzymes by studying inactivation of GPx1 and Prx II in human RBCs. Mass spectrometry revealed that treatment of GPx1 with H2O2 converts the selenocysteine residue at its active site to dehydroalanine (DHA). We developed a blot method for detection of DHA-containing proteins, with which we observed that the amount of DHA-containing GPx1 increases with increasing RBC density, which is correlated with increasing RBC age. Given that the conversion of selenocysteine to DHA is irreversible, the content of DHA-GPx1 in each RBC likely reflects total oxidative stress experienced by the cell during its lifetime. Prx II is inactivated by occasional hyperoxidation of its catalytic cysteine to cysteine sulfinic acid during catalysis. We believe that the activity of sulfiredoxin in RBCs is sufficient to counteract the hyperoxidation of Prx II that occurs in the presence of the basal level of H2O2 flux resulting from hemoglobin autoxidation. If the H2O2 flux is increased above the basal level, however, the sulfinic Prx II begins to accumulate. In the presence of an increased H2O2 flux, inhibition of catalase accelerated the accumulation of sulfinic Prx II, indicative of the protective role of catalase. Antioxid. Redox Signal. 12, 1235-1246.

AB - Catalase, glutathione peroxidase1 (GPx1), and peroxiredoxin (Prx) II are the principal enzymes responsible for peroxide elimination in RBC. We have now evaluated the relative roles of these enzymes by studying inactivation of GPx1 and Prx II in human RBCs. Mass spectrometry revealed that treatment of GPx1 with H2O2 converts the selenocysteine residue at its active site to dehydroalanine (DHA). We developed a blot method for detection of DHA-containing proteins, with which we observed that the amount of DHA-containing GPx1 increases with increasing RBC density, which is correlated with increasing RBC age. Given that the conversion of selenocysteine to DHA is irreversible, the content of DHA-GPx1 in each RBC likely reflects total oxidative stress experienced by the cell during its lifetime. Prx II is inactivated by occasional hyperoxidation of its catalytic cysteine to cysteine sulfinic acid during catalysis. We believe that the activity of sulfiredoxin in RBCs is sufficient to counteract the hyperoxidation of Prx II that occurs in the presence of the basal level of H2O2 flux resulting from hemoglobin autoxidation. If the H2O2 flux is increased above the basal level, however, the sulfinic Prx II begins to accumulate. In the presence of an increased H2O2 flux, inhibition of catalase accelerated the accumulation of sulfinic Prx II, indicative of the protective role of catalase. Antioxid. Redox Signal. 12, 1235-1246.

UR - http://www.scopus.com/inward/record.url?scp=77951813369&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77951813369&partnerID=8YFLogxK

U2 - 10.1089/ars.2009.2701

DO - 10.1089/ars.2009.2701

M3 - Article

C2 - 20070187

AN - SCOPUS:77951813369

VL - 12

SP - 1235

EP - 1246

JO - Antioxidants and Redox Signaling

JF - Antioxidants and Redox Signaling

SN - 1523-0864

IS - 11

ER -