Removal of amino acid, peptide and protein hydroperoxides by reaction with peroxiredoxins 2 and 3
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Removal of amino acid, peptide and protein hydroperoxides by reaction with peroxiredoxins 2 and 3. / Peskin, Alexander V; Cox, Andrew G; Nagy, Péter; Morgan, Philip E; Hampton, Mark B; Davies, Michael Jonathan; Winterbourn, Christine C.
In: Biochemical Journal, Vol. 432, No. 2, 01.12.2010, p. 313-21.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Removal of amino acid, peptide and protein hydroperoxides by reaction with peroxiredoxins 2 and 3
AU - Peskin, Alexander V
AU - Cox, Andrew G
AU - Nagy, Péter
AU - Morgan, Philip E
AU - Hampton, Mark B
AU - Davies, Michael Jonathan
AU - Winterbourn, Christine C
PY - 2010/12/1
Y1 - 2010/12/1
N2 - Prxs (peroxiredoxins) are a ubiquitous family of cysteine-dependent peroxidases that react rapidly with H2O2 and alkyl hydroperoxides and provide defence against these reactive oxidants. Hydroperoxides are also formed on amino acids and proteins during oxidative stress, and they too are a potential cause of biological damage. We have investigated whether Prxs react with amino acid, peptide and protein hydroperoxides, and whether the reactions are sufficiently rapid for these enzymes to provide antioxidant protection against these oxidants. Isolated Prx2, which is a cytosolic protein, and Prx3, which resides within mitochondria, were reacted with a selection of hydroperoxides generated by γ-radiolysis or singlet oxygen, on free amino acids, peptides and proteins. Reactions were followed by measuring the accumulation of disulfide-linked Prx dimers, via non-reducing SDS/PAGE, or the loss of the corresponding hydroperoxide, using quench-flow and LC (liquid chromatography)/MS. All the hydroperoxides induced rapid oxidation, with little difference in reactivity between Prx2 and Prx3. N-acetyl leucine hydroperoxides reacted with Prx2 with a rate constant of 4 × 10(4) M-1 · s-1. Hydroperoxides present on leucine, isoleucine or tyrosine reacted at a comparable rate, whereas histidine hydroperoxides were ~10-fold less reactive. Hydroperoxides present on lysozyme and BSA reacted with rate constants of ~100 M-1 · s-1. Addition of an uncharged derivative of leucine hydroperoxide to intact erythrocytes caused Prx2 oxidation with no concomitant loss in GSH, as did BSA hydroperoxide when added to concentrated erythrocyte lysate. Prxs are therefore favoured intracellular targets for peptide/protein hydroperoxides and have the potential to detoxify these species in vivo.
AB - Prxs (peroxiredoxins) are a ubiquitous family of cysteine-dependent peroxidases that react rapidly with H2O2 and alkyl hydroperoxides and provide defence against these reactive oxidants. Hydroperoxides are also formed on amino acids and proteins during oxidative stress, and they too are a potential cause of biological damage. We have investigated whether Prxs react with amino acid, peptide and protein hydroperoxides, and whether the reactions are sufficiently rapid for these enzymes to provide antioxidant protection against these oxidants. Isolated Prx2, which is a cytosolic protein, and Prx3, which resides within mitochondria, were reacted with a selection of hydroperoxides generated by γ-radiolysis or singlet oxygen, on free amino acids, peptides and proteins. Reactions were followed by measuring the accumulation of disulfide-linked Prx dimers, via non-reducing SDS/PAGE, or the loss of the corresponding hydroperoxide, using quench-flow and LC (liquid chromatography)/MS. All the hydroperoxides induced rapid oxidation, with little difference in reactivity between Prx2 and Prx3. N-acetyl leucine hydroperoxides reacted with Prx2 with a rate constant of 4 × 10(4) M-1 · s-1. Hydroperoxides present on leucine, isoleucine or tyrosine reacted at a comparable rate, whereas histidine hydroperoxides were ~10-fold less reactive. Hydroperoxides present on lysozyme and BSA reacted with rate constants of ~100 M-1 · s-1. Addition of an uncharged derivative of leucine hydroperoxide to intact erythrocytes caused Prx2 oxidation with no concomitant loss in GSH, as did BSA hydroperoxide when added to concentrated erythrocyte lysate. Prxs are therefore favoured intracellular targets for peptide/protein hydroperoxides and have the potential to detoxify these species in vivo.
KW - Amino Acids
KW - Dipeptides
KW - Erythrocytes
KW - Gamma Rays
KW - Glutathione
KW - Humans
KW - Hydrogen Peroxide
KW - Isoleucine
KW - Leucine
KW - Oxidative Stress
KW - Peptides
KW - Peroxides
KW - Peroxiredoxin III
KW - Peroxiredoxins
KW - Singlet Oxygen
U2 - 10.1042/BJ20101156
DO - 10.1042/BJ20101156
M3 - Journal article
C2 - 20840079
VL - 432
SP - 313
EP - 321
JO - Biochemical Journal
JF - Biochemical Journal
SN - 0264-6021
IS - 2
ER -
ID: 129669928