Nitrotyrosine as a new marker for endogenous nitrosation and nitration of proteins

H. Ohshima, M. Friesen, I. Brouet, H. Bartsch

Research output: Contribution to journalArticle

Abstract

3-Nitrotyrosine (NTYR) in tissue or blood proteins was evaluated as a possible exposure marker for exogenous and endogenous nitrosating or nitrating agents. A sensitive and selective method for analysing NTYR by gas chromatography with a thermal energy analyser (GC-TEA) was developed. Using this method, a number of kinetic studies were carried out. It was found that free and protein-bound tyrosine residues easily react with nitrating/nitrosating agents to yield NTYR. NTYR formation in vivo showed a dose-dependent increase in NTYR in both plasma proteins and haemoglobin obtained from rats 24 hr after ip injection of various doses (0.5-2.5 μmol/rat) of tetranitromethane. Major urinary metabolites of NTYR, given orally to rats, were isolated and identified as 3-nitro-4-hydroxyphenylacetic acid (NHPA) and 3-nitro-4-hydroxyphenyllactic acid (NHPL). About 44% and 5% of the oral dose of NTYR (100 μg/rat) was excreted as NHPA and NHPL, respectively. Eleven 24-hr human urine samples were analysed for NHPA by GC-TEA after ethyl acetate extraction and HPLC purification: quantities ranging from 0 to 7.9 μg/24 hr, mean ± SD 2.8 ± 2.3 (n = 11) were detected (detection limit 0.2 μg/litre). NTYR in proteins or its metabolites in urine can be readily analysed by GC-TEA as a new/additional marker for endogenous nitrosation and nitration.

Original languageEnglish (US)
Pages (from-to)647-652
Number of pages6
JournalFood and Chemical Toxicology
Volume28
Issue number9
DOIs
StatePublished - 1990
Externally publishedYes

Fingerprint

Nitrosation
Nitration
acids
gas chromatography
rats
Rats
Proteins
heat
proteins
Thermal energy
blood proteins
Gas chromatography
Gas Chromatography
energy
urine
dosage
Hot Temperature
metabolites
Metabolites
Blood Proteins

Keywords

  • 3-nitro-4-hydroxyphenylacetic acid
  • 3-nitro-4-hydroxyphenyllactic acid
  • 3-nitrotyrosine
  • bovine serum albumin
  • BSA
  • gas chromatography with a thermal energy analyser
  • gas chromatography-mass spectrometry
  • GC-MS
  • GC-TEA
  • MTBSTFA
  • N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide
  • NHPA
  • NHPL
  • nitrosoproline
  • NPRO
  • NTYR
  • tetranitromethane
  • TNM

ASJC Scopus subject areas

  • Food Science
  • Toxicology

Cite this

Nitrotyrosine as a new marker for endogenous nitrosation and nitration of proteins. / Ohshima, H.; Friesen, M.; Brouet, I.; Bartsch, H.

In: Food and Chemical Toxicology, Vol. 28, No. 9, 1990, p. 647-652.

Research output: Contribution to journalArticle

Ohshima, H. ; Friesen, M. ; Brouet, I. ; Bartsch, H. / Nitrotyrosine as a new marker for endogenous nitrosation and nitration of proteins. In: Food and Chemical Toxicology. 1990 ; Vol. 28, No. 9. pp. 647-652.
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T1 - Nitrotyrosine as a new marker for endogenous nitrosation and nitration of proteins

AU - Ohshima, H.

AU - Friesen, M.

AU - Brouet, I.

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PY - 1990

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N2 - 3-Nitrotyrosine (NTYR) in tissue or blood proteins was evaluated as a possible exposure marker for exogenous and endogenous nitrosating or nitrating agents. A sensitive and selective method for analysing NTYR by gas chromatography with a thermal energy analyser (GC-TEA) was developed. Using this method, a number of kinetic studies were carried out. It was found that free and protein-bound tyrosine residues easily react with nitrating/nitrosating agents to yield NTYR. NTYR formation in vivo showed a dose-dependent increase in NTYR in both plasma proteins and haemoglobin obtained from rats 24 hr after ip injection of various doses (0.5-2.5 μmol/rat) of tetranitromethane. Major urinary metabolites of NTYR, given orally to rats, were isolated and identified as 3-nitro-4-hydroxyphenylacetic acid (NHPA) and 3-nitro-4-hydroxyphenyllactic acid (NHPL). About 44% and 5% of the oral dose of NTYR (100 μg/rat) was excreted as NHPA and NHPL, respectively. Eleven 24-hr human urine samples were analysed for NHPA by GC-TEA after ethyl acetate extraction and HPLC purification: quantities ranging from 0 to 7.9 μg/24 hr, mean ± SD 2.8 ± 2.3 (n = 11) were detected (detection limit 0.2 μg/litre). NTYR in proteins or its metabolites in urine can be readily analysed by GC-TEA as a new/additional marker for endogenous nitrosation and nitration.

AB - 3-Nitrotyrosine (NTYR) in tissue or blood proteins was evaluated as a possible exposure marker for exogenous and endogenous nitrosating or nitrating agents. A sensitive and selective method for analysing NTYR by gas chromatography with a thermal energy analyser (GC-TEA) was developed. Using this method, a number of kinetic studies were carried out. It was found that free and protein-bound tyrosine residues easily react with nitrating/nitrosating agents to yield NTYR. NTYR formation in vivo showed a dose-dependent increase in NTYR in both plasma proteins and haemoglobin obtained from rats 24 hr after ip injection of various doses (0.5-2.5 μmol/rat) of tetranitromethane. Major urinary metabolites of NTYR, given orally to rats, were isolated and identified as 3-nitro-4-hydroxyphenylacetic acid (NHPA) and 3-nitro-4-hydroxyphenyllactic acid (NHPL). About 44% and 5% of the oral dose of NTYR (100 μg/rat) was excreted as NHPA and NHPL, respectively. Eleven 24-hr human urine samples were analysed for NHPA by GC-TEA after ethyl acetate extraction and HPLC purification: quantities ranging from 0 to 7.9 μg/24 hr, mean ± SD 2.8 ± 2.3 (n = 11) were detected (detection limit 0.2 μg/litre). NTYR in proteins or its metabolites in urine can be readily analysed by GC-TEA as a new/additional marker for endogenous nitrosation and nitration.

KW - 3-nitro-4-hydroxyphenylacetic acid

KW - 3-nitro-4-hydroxyphenyllactic acid

KW - 3-nitrotyrosine

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KW - BSA

KW - gas chromatography with a thermal energy analyser

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KW - GC-TEA

KW - MTBSTFA

KW - N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide

KW - NHPA

KW - NHPL

KW - nitrosoproline

KW - NPRO

KW - NTYR

KW - tetranitromethane

KW - TNM

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