A tale of two controversies. Defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase-deficient mice, and the nature of peroxidase-generated reactive nitrogen species

Marie Luise Brennan, Weijia Wu, Xiaoming Fu, Zhongzhu Shen, Wei Song, Heather Frost, Caryn Vadseth, Laura Narine, Elizabeth Lenkiewicz, Michael T. Borchers, Aldons J. Lusis, James J. Lee, Nancy A Lee, Husam M. Abu-Soud, Harry Ischiropoulos, Stanley L. Hazen

Research output: Contribution to journalArticle

423 Citations (Scopus)

Abstract

Nitrotyrosine is widely used as a marker of post-translational modification by the nitric oxide (.NO, nitrogen monoxide)-derived oxidant peroxynitrite (ONOO-). However, since the discovery that myeloperoxidase (MPO) and eosinophil peroxidase (EPO) can generate nitrotyrosine via oxidation of nitrite (NO2 -), several questions have arisen. First, the relative contribution of peroxidases to nitrotyrosine formation in vivo is unknown. Further, although evidence suggests that the one-electron oxidation product, nitrogen dioxide (.NO2), is the primary species formed, neither a direct demonstration that peroxidases form this gas nor studies designed to test for the possible concomitant formation of the two-electron oxidation product, ONOO-, have been reported. Using multiple distinct models of acute inflammation with EPO- and MPO-knockout mice, we now demonstrate that leukocyte peroxidases participate in nitrotyrosine formation in vivo. In some models, MPO and EPO played a dominant role, accounting for the majority of nitrotyrosine formed. However, in other leukocyte-rich acute inflammatory models, no contribution for either MPO or EPO to nitrotyrosine formation could be demonstrated. Head-space gas analysis of heliumswept reaction mixtures provides direct evidence that leukocyte peroxidases catalytically generate .NO2 formation using H2O2 and NO2 - as substrates. However, formation of an additional oxidant was suggested since both enzymes promote NO2 --dependent hydroxylation of targets under acidic conditions, a chemical reactivity shared with ONOO- but not .NO2. Collectively, our results demonstrate that: 1) MPO and EPO contribute to tyrosine nitration in vivo; 2) the major reactive nitrogen species formed by leukocyte peroxidase-catalyzed oxidation of NO2 - is the one-electron oxidation product, ̇NO2; 3) as a minor reaction, peroxidases may also catalyze the two-electron oxidation of NO2 -, producing a ONOO--like product. We speculate that the latter reaction generates a labile Fe-ONOO complex, which may be released following protonation under acidic conditions such as might exist at sites of inflammation.

Original languageEnglish (US)
Pages (from-to)17415-17427
Number of pages13
JournalJournal of Biological Chemistry
Volume277
Issue number20
DOIs
StatePublished - May 17 2002

Fingerprint

Eosinophil Peroxidase
Reactive Nitrogen Species
Peroxidases
Peroxidase
Oxidation
Leukocytes
Electrons
Oxidants
Nitric Oxide
Gases
Inflammation
Nitration
Nitrogen Dioxide
Chemical reactivity
Hydroxylation
Peroxynitrous Acid
Gas fuel analysis
Protonation
Post Translational Protein Processing
Nitrites

ASJC Scopus subject areas

  • Biochemistry

Cite this

A tale of two controversies. Defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase-deficient mice, and the nature of peroxidase-generated reactive nitrogen species. / Brennan, Marie Luise; Wu, Weijia; Fu, Xiaoming; Shen, Zhongzhu; Song, Wei; Frost, Heather; Vadseth, Caryn; Narine, Laura; Lenkiewicz, Elizabeth; Borchers, Michael T.; Lusis, Aldons J.; Lee, James J.; Lee, Nancy A; Abu-Soud, Husam M.; Ischiropoulos, Harry; Hazen, Stanley L.

In: Journal of Biological Chemistry, Vol. 277, No. 20, 17.05.2002, p. 17415-17427.

Research output: Contribution to journalArticle

Brennan, ML, Wu, W, Fu, X, Shen, Z, Song, W, Frost, H, Vadseth, C, Narine, L, Lenkiewicz, E, Borchers, MT, Lusis, AJ, Lee, JJ, Lee, NA, Abu-Soud, HM, Ischiropoulos, H & Hazen, SL 2002, 'A tale of two controversies. Defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase-deficient mice, and the nature of peroxidase-generated reactive nitrogen species', Journal of Biological Chemistry, vol. 277, no. 20, pp. 17415-17427. https://doi.org/10.1074/jbc.M112400200
Brennan, Marie Luise ; Wu, Weijia ; Fu, Xiaoming ; Shen, Zhongzhu ; Song, Wei ; Frost, Heather ; Vadseth, Caryn ; Narine, Laura ; Lenkiewicz, Elizabeth ; Borchers, Michael T. ; Lusis, Aldons J. ; Lee, James J. ; Lee, Nancy A ; Abu-Soud, Husam M. ; Ischiropoulos, Harry ; Hazen, Stanley L. / A tale of two controversies. Defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase-deficient mice, and the nature of peroxidase-generated reactive nitrogen species. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 20. pp. 17415-17427.
@article{931c692e0faa49d0b3436024db06eaa3,
title = "A tale of two controversies. Defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase-deficient mice, and the nature of peroxidase-generated reactive nitrogen species",
abstract = "Nitrotyrosine is widely used as a marker of post-translational modification by the nitric oxide (.NO, nitrogen monoxide)-derived oxidant peroxynitrite (ONOO-). However, since the discovery that myeloperoxidase (MPO) and eosinophil peroxidase (EPO) can generate nitrotyrosine via oxidation of nitrite (NO2 -), several questions have arisen. First, the relative contribution of peroxidases to nitrotyrosine formation in vivo is unknown. Further, although evidence suggests that the one-electron oxidation product, nitrogen dioxide (.NO2), is the primary species formed, neither a direct demonstration that peroxidases form this gas nor studies designed to test for the possible concomitant formation of the two-electron oxidation product, ONOO-, have been reported. Using multiple distinct models of acute inflammation with EPO- and MPO-knockout mice, we now demonstrate that leukocyte peroxidases participate in nitrotyrosine formation in vivo. In some models, MPO and EPO played a dominant role, accounting for the majority of nitrotyrosine formed. However, in other leukocyte-rich acute inflammatory models, no contribution for either MPO or EPO to nitrotyrosine formation could be demonstrated. Head-space gas analysis of heliumswept reaction mixtures provides direct evidence that leukocyte peroxidases catalytically generate .NO2 formation using H2O2 and NO2 - as substrates. However, formation of an additional oxidant was suggested since both enzymes promote NO2 --dependent hydroxylation of targets under acidic conditions, a chemical reactivity shared with ONOO- but not .NO2. Collectively, our results demonstrate that: 1) MPO and EPO contribute to tyrosine nitration in vivo; 2) the major reactive nitrogen species formed by leukocyte peroxidase-catalyzed oxidation of NO2 - is the one-electron oxidation product, ̇NO2; 3) as a minor reaction, peroxidases may also catalyze the two-electron oxidation of NO2 -, producing a ONOO--like product. We speculate that the latter reaction generates a labile Fe-ONOO complex, which may be released following protonation under acidic conditions such as might exist at sites of inflammation.",
author = "Brennan, {Marie Luise} and Weijia Wu and Xiaoming Fu and Zhongzhu Shen and Wei Song and Heather Frost and Caryn Vadseth and Laura Narine and Elizabeth Lenkiewicz and Borchers, {Michael T.} and Lusis, {Aldons J.} and Lee, {James J.} and Lee, {Nancy A} and Abu-Soud, {Husam M.} and Harry Ischiropoulos and Hazen, {Stanley L.}",
year = "2002",
month = "5",
day = "17",
doi = "10.1074/jbc.M112400200",
language = "English (US)",
volume = "277",
pages = "17415--17427",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "20",

}

TY - JOUR

T1 - A tale of two controversies. Defining both the role of peroxidases in nitrotyrosine formation in vivo using eosinophil peroxidase and myeloperoxidase-deficient mice, and the nature of peroxidase-generated reactive nitrogen species

AU - Brennan, Marie Luise

AU - Wu, Weijia

AU - Fu, Xiaoming

AU - Shen, Zhongzhu

AU - Song, Wei

AU - Frost, Heather

AU - Vadseth, Caryn

AU - Narine, Laura

AU - Lenkiewicz, Elizabeth

AU - Borchers, Michael T.

AU - Lusis, Aldons J.

AU - Lee, James J.

AU - Lee, Nancy A

AU - Abu-Soud, Husam M.

AU - Ischiropoulos, Harry

AU - Hazen, Stanley L.

PY - 2002/5/17

Y1 - 2002/5/17

N2 - Nitrotyrosine is widely used as a marker of post-translational modification by the nitric oxide (.NO, nitrogen monoxide)-derived oxidant peroxynitrite (ONOO-). However, since the discovery that myeloperoxidase (MPO) and eosinophil peroxidase (EPO) can generate nitrotyrosine via oxidation of nitrite (NO2 -), several questions have arisen. First, the relative contribution of peroxidases to nitrotyrosine formation in vivo is unknown. Further, although evidence suggests that the one-electron oxidation product, nitrogen dioxide (.NO2), is the primary species formed, neither a direct demonstration that peroxidases form this gas nor studies designed to test for the possible concomitant formation of the two-electron oxidation product, ONOO-, have been reported. Using multiple distinct models of acute inflammation with EPO- and MPO-knockout mice, we now demonstrate that leukocyte peroxidases participate in nitrotyrosine formation in vivo. In some models, MPO and EPO played a dominant role, accounting for the majority of nitrotyrosine formed. However, in other leukocyte-rich acute inflammatory models, no contribution for either MPO or EPO to nitrotyrosine formation could be demonstrated. Head-space gas analysis of heliumswept reaction mixtures provides direct evidence that leukocyte peroxidases catalytically generate .NO2 formation using H2O2 and NO2 - as substrates. However, formation of an additional oxidant was suggested since both enzymes promote NO2 --dependent hydroxylation of targets under acidic conditions, a chemical reactivity shared with ONOO- but not .NO2. Collectively, our results demonstrate that: 1) MPO and EPO contribute to tyrosine nitration in vivo; 2) the major reactive nitrogen species formed by leukocyte peroxidase-catalyzed oxidation of NO2 - is the one-electron oxidation product, ̇NO2; 3) as a minor reaction, peroxidases may also catalyze the two-electron oxidation of NO2 -, producing a ONOO--like product. We speculate that the latter reaction generates a labile Fe-ONOO complex, which may be released following protonation under acidic conditions such as might exist at sites of inflammation.

AB - Nitrotyrosine is widely used as a marker of post-translational modification by the nitric oxide (.NO, nitrogen monoxide)-derived oxidant peroxynitrite (ONOO-). However, since the discovery that myeloperoxidase (MPO) and eosinophil peroxidase (EPO) can generate nitrotyrosine via oxidation of nitrite (NO2 -), several questions have arisen. First, the relative contribution of peroxidases to nitrotyrosine formation in vivo is unknown. Further, although evidence suggests that the one-electron oxidation product, nitrogen dioxide (.NO2), is the primary species formed, neither a direct demonstration that peroxidases form this gas nor studies designed to test for the possible concomitant formation of the two-electron oxidation product, ONOO-, have been reported. Using multiple distinct models of acute inflammation with EPO- and MPO-knockout mice, we now demonstrate that leukocyte peroxidases participate in nitrotyrosine formation in vivo. In some models, MPO and EPO played a dominant role, accounting for the majority of nitrotyrosine formed. However, in other leukocyte-rich acute inflammatory models, no contribution for either MPO or EPO to nitrotyrosine formation could be demonstrated. Head-space gas analysis of heliumswept reaction mixtures provides direct evidence that leukocyte peroxidases catalytically generate .NO2 formation using H2O2 and NO2 - as substrates. However, formation of an additional oxidant was suggested since both enzymes promote NO2 --dependent hydroxylation of targets under acidic conditions, a chemical reactivity shared with ONOO- but not .NO2. Collectively, our results demonstrate that: 1) MPO and EPO contribute to tyrosine nitration in vivo; 2) the major reactive nitrogen species formed by leukocyte peroxidase-catalyzed oxidation of NO2 - is the one-electron oxidation product, ̇NO2; 3) as a minor reaction, peroxidases may also catalyze the two-electron oxidation of NO2 -, producing a ONOO--like product. We speculate that the latter reaction generates a labile Fe-ONOO complex, which may be released following protonation under acidic conditions such as might exist at sites of inflammation.

UR - http://www.scopus.com/inward/record.url?scp=0037124020&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037124020&partnerID=8YFLogxK

U2 - 10.1074/jbc.M112400200

DO - 10.1074/jbc.M112400200

M3 - Article

VL - 277

SP - 17415

EP - 17427

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 20

ER -