Nitration of unsaturated fatty acids by nitric oxide-derived reactive nitrogen species peroxynitrite, nitrous acid, nitrogen dioxide, and nitronium ion

Valerie B. O'Donnell, Jason P. Eiserich, Phillip H. Chumley, Michael J. Jablonsky, N. Rama Krishna, Marion Kirk, Stephen Barnes, Victor M. Darley-Usmar, Bruce A. Freeman

Research output: Contribution to journalArticle

216 Citations (Scopus)

Abstract

Reactive nitrogen species derived from nitric oxide are potent oxidants formed during inflammation that can oxidize membrane and lipoprotein lipids in vivo. Herein, it is demonstrated that several of these species react with unsaturated fatty acid to yield nitrated oxidation products. Using HPLC coupled with both UV detection and electrospray ionization mass spectrometry, products of reaction of ONOO- with linoleic acid displayed mass/charge (m/z) characteristics of LNO2 (at least three products at m/z 324, negative ion mode). Further analysis by MS/MS gave a major fragment at m/z 46. Addition of a NO2 group was confirmed using [15N]ONOO- which gave a product at m/z 325, fragmenting to form a daughter ion at m/z 47. Formation of nitrated lipids was inhibited by bicarbonate, superoxide dismutase (SOD), and Fe3+- EDTA, while the yield of oxidation products was decreased by bicarbonate and SOD, but not by Fe3+-EDTA. Reaction of linoleic acid with both nitrogen dioxide (·NO2) or nitronium tetrafluoroborate (NO2BF4) also yielded nitrated lipid products (m/z 324), with HPLC retention times and MS/MS fragmentation patterns identical to the m/z 324 species formed by reaction of ONOO- with linoleic acid. Finally, reaction of HPODE, but not linoleate, with nitrous acid (HONO) or isobutyl nitrite (Bu(i)ONO) yielded a product at m/z 340, or 341 upon reacting with [15N]HONO. MS/MS analysis gave an NO2 - fragment, and 15N NMR indicated that the product contained a nitro (RNO2) functional group, suggesting that the product was nitroepoxylinoleic acid [L(O)NO2]. This species could form via homolytic dissociation of LOONO to LO· and ·NO2 and rearrangement of LO· to an epoxyallylic radical L(O)· followed by recombination of L(O)· with ·NO2. Since unsaturated lipids of membranes and lipoproteins are critical targets of reactive oxygen and nitrogen species, these pathways lend insight into mechanisms for the formation of novel nitrogen-containing lipid products in vivo and provide synthetic strategies for further structural and functional studies.

Original languageEnglish (US)
Pages (from-to)83-92
Number of pages10
JournalChemical Research in Toxicology
Volume12
Issue number1
DOIs
StatePublished - Jan 1999
Externally publishedYes

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Nitrous Acid
Nitration
Reactive Nitrogen Species
Nitrogen Dioxide
Peroxynitrous Acid
Linoleic Acid
Unsaturated Fatty Acids
Nitric Oxide
Ions
Lipids
Membrane Lipids
Bicarbonates
Edetic Acid
Lipoproteins
Superoxide Dismutase
High Pressure Liquid Chromatography
Electrospray ionization
Oxidation
Electrospray Ionization Mass Spectrometry
Oxidants

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Chemistry(all)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Nitration of unsaturated fatty acids by nitric oxide-derived reactive nitrogen species peroxynitrite, nitrous acid, nitrogen dioxide, and nitronium ion. / O'Donnell, Valerie B.; Eiserich, Jason P.; Chumley, Phillip H.; Jablonsky, Michael J.; Krishna, N. Rama; Kirk, Marion; Barnes, Stephen; Darley-Usmar, Victor M.; Freeman, Bruce A.

In: Chemical Research in Toxicology, Vol. 12, No. 1, 01.1999, p. 83-92.

Research output: Contribution to journalArticle

O'Donnell, VB, Eiserich, JP, Chumley, PH, Jablonsky, MJ, Krishna, NR, Kirk, M, Barnes, S, Darley-Usmar, VM & Freeman, BA 1999, 'Nitration of unsaturated fatty acids by nitric oxide-derived reactive nitrogen species peroxynitrite, nitrous acid, nitrogen dioxide, and nitronium ion', Chemical Research in Toxicology, vol. 12, no. 1, pp. 83-92. https://doi.org/10.1021/tx980207u
O'Donnell, Valerie B. ; Eiserich, Jason P. ; Chumley, Phillip H. ; Jablonsky, Michael J. ; Krishna, N. Rama ; Kirk, Marion ; Barnes, Stephen ; Darley-Usmar, Victor M. ; Freeman, Bruce A. / Nitration of unsaturated fatty acids by nitric oxide-derived reactive nitrogen species peroxynitrite, nitrous acid, nitrogen dioxide, and nitronium ion. In: Chemical Research in Toxicology. 1999 ; Vol. 12, No. 1. pp. 83-92.
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AU - Eiserich, Jason P.

AU - Chumley, Phillip H.

AU - Jablonsky, Michael J.

AU - Krishna, N. Rama

AU - Kirk, Marion

AU - Barnes, Stephen

AU - Darley-Usmar, Victor M.

AU - Freeman, Bruce A.

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N2 - Reactive nitrogen species derived from nitric oxide are potent oxidants formed during inflammation that can oxidize membrane and lipoprotein lipids in vivo. Herein, it is demonstrated that several of these species react with unsaturated fatty acid to yield nitrated oxidation products. Using HPLC coupled with both UV detection and electrospray ionization mass spectrometry, products of reaction of ONOO- with linoleic acid displayed mass/charge (m/z) characteristics of LNO2 (at least three products at m/z 324, negative ion mode). Further analysis by MS/MS gave a major fragment at m/z 46. Addition of a NO2 group was confirmed using [15N]ONOO- which gave a product at m/z 325, fragmenting to form a daughter ion at m/z 47. Formation of nitrated lipids was inhibited by bicarbonate, superoxide dismutase (SOD), and Fe3+- EDTA, while the yield of oxidation products was decreased by bicarbonate and SOD, but not by Fe3+-EDTA. Reaction of linoleic acid with both nitrogen dioxide (·NO2) or nitronium tetrafluoroborate (NO2BF4) also yielded nitrated lipid products (m/z 324), with HPLC retention times and MS/MS fragmentation patterns identical to the m/z 324 species formed by reaction of ONOO- with linoleic acid. Finally, reaction of HPODE, but not linoleate, with nitrous acid (HONO) or isobutyl nitrite (Bu(i)ONO) yielded a product at m/z 340, or 341 upon reacting with [15N]HONO. MS/MS analysis gave an NO2 - fragment, and 15N NMR indicated that the product contained a nitro (RNO2) functional group, suggesting that the product was nitroepoxylinoleic acid [L(O)NO2]. This species could form via homolytic dissociation of LOONO to LO· and ·NO2 and rearrangement of LO· to an epoxyallylic radical L(O)· followed by recombination of L(O)· with ·NO2. Since unsaturated lipids of membranes and lipoproteins are critical targets of reactive oxygen and nitrogen species, these pathways lend insight into mechanisms for the formation of novel nitrogen-containing lipid products in vivo and provide synthetic strategies for further structural and functional studies.

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