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Site-specific protein adducts of 4-hydroxy-2(E)-nonenal in human THP-1 monocytic cells: protein carbonylation is

Juan Chavez1, Woon-Gye Chung, Cristobal L Miranda

  • 1Department of Chemistry, Oregon State University, Corvallis, Oregon 97331, USA.

Chemical Research in Toxicology
|January 2, 2010
PubMed

Insights

This study identified specific proteins and amino acid sites modified by 4-hydroxy-2(E)-nonenal (HNE) in human cells. Ascorbic acid reduced these HNE-protein adducts, highlighting potential therapeutic avenues.

Area of Science:

  • Biochemistry
  • Proteomics
  • Cell Biology

Background:

  • 4-hydroxy-2(E)-nonenal (HNE) is a reactive aldehyde implicated in cellular damage.
  • Understanding HNE's protein targets is crucial for elucidating its role in disease.

Purpose of the Study:

  • To identify specific protein targets and modification sites of HNE in human monocytic THP-1 cells.
  • To investigate the protective effect of ascorbic acid against HNE-induced protein modification.

Main Methods:

  • Utilized a multipronged proteomic approach including electrophoresis, immunoblotting, and mass spectrometry (LC-MS/MS).
  • Employed biotinylation with an aldehyde-reactive probe (ARP) for enrichment and identification of HNE-protein adducts.
  • Analyzed modifications at the site-specific level for identified proteins.

Main Results:

  • Identified 16 proteins as targets of HNE adduction.
  • Assigned 18 specific modification sites, primarily at Cysteine (Cys) and Histidine (His) residues.
  • Observed HNE modification of proteins involved in cytoskeleton, stress response, and metabolic pathways, including novel site-specific adducts.

Conclusions:

  • Provides the first evidence of site-specific HNE adduction to key proteins like tubulin, alpha-actinin-4, vimentin, D-3-phosphoglycerate dehydrogenase, and aldolase A.
  • Demonstrates that ascorbic acid can reduce HNE-protein adduct formation.
  • Advances the understanding of HNE-mediated cellular damage and potential mitigation strategies.

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