Localization of 4-hydroxy-2-nonenal-modified proteins in kidney following iron overload

Theodor A. Zainal, Richard Weindruch, Luke I. Szweda, Terry D. Oberley

Research output: Contribution to journalArticlepeer-review

47 Scopus citations


Intraperitoneal (IP) injection of ferric nitrilotriacetate (Fe-NTA) to rats and mice results in iron-induced free radical injury and cancer in kidneys. We sought to clarify the exact localization of acute oxidative damage in Fe-NTA-induced nephrotoxicity by performing immunogold light and electron microscopic (EM) techniques using an antibody against 4-hydroxy-2- nonenal (HNE)-modified proteins. Biochemical assays were done to provide complementary quantitative data. Renal accumulation of lipid peroxidation- derived aldehydes, such as malondialdehyde (MDA) and 4-hydroxy-2-alkenals (4- HDA), increased in parallel with protein carbonyl content, an indicator of protein oxidation, 30 min after administration of Fe-NTA. Immunogold light microscopy showed that HNE-modified proteins increased at 30 min with positivity localized to proximal tubular cells. Immunogold EM demonstrated that HNE-modified proteins were mainly in the mitochondria and nuclei of the proximal tubular epithelium. The intensity of labeling at both the light and EM levels increased together with levels of biochemically measured lipid peroxidation products and protein carbonyl content. Our data suggest that the mechanism of acute nephrotoxicity of Fe-NTA involves mitochondrial and nuclear oxidative damage, findings that may help to define the mechanisms of iron-induced cell injury.

Original languageEnglish (US)
Pages (from-to)1181-1193
Number of pages13
JournalFree Radical Biology and Medicine
Issue number9-10
StatePublished - May 1999


  • 4-Hydroxy-2-nonenal
  • Aldehydes
  • Ferric nitrilotriacetate
  • Free radicals
  • Immunogold
  • Iron
  • Lipid peroxidation
  • Reactive oxygen species

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)


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