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Inactivation of NADP+-dependent isocitrate dehydrogenase by lipid peroxidation products

Joon-Hyuck Yang1, Eun Sun Yang, Jeen-Woo Park

  • 1Department of Biochemistry, College of Natural Sciences, Kyungpook National University, Taegu 702-701, South Korea.

Free Radical Research
|May 8, 2004
PubMed

Insights

NADP+-dependent isocitrate dehydrogenase (ICDH) protects cells from oxidative damage by supplying NADPH. Lipid peroxidation products damage ICDH, impairing antioxidant defenses and potentially causing a prooxidant state.

Area of Science:

  • Biochemistry
  • Cellular Redox Biology
  • Oxidative Stress

Background:

  • Membrane lipid peroxidation generates reactive products that can modify proteins.
  • NADP+-dependent isocitrate dehydrogenase (ICDH) plays a crucial role in maintaining cellular redox balance by supplying NADPH for antioxidant systems.

Purpose of the Study:

  • To investigate the susceptibility of ICDH to oxidative damage induced by lipid peroxidation products.
  • To elucidate the functional and structural consequences of ICDH modification by lipid peroxidation.

Main Methods:

  • Enzyme activity assays
  • Spectroscopic techniques (thermal stability, fluorescence)
  • Hydrophobic probe binding assays
  • Cellular treatment with lipid peroxidation inducers (AAPH)
  • Immunoprecipitation and immunoblotting to detect adducts

Main Results:

  • ICDH activity decreased and carbonyl groups formed upon exposure to lipid peroxidation products (MDA, HNE, lipid hydroperoxide).
  • Structural changes in ICDH included altered thermal stability and fluorescence properties.
  • AAPH treatment significantly reduced both cytosolic and mitochondrial ICDH activity in U937 cells.
  • 4-hydroxynonenal (HNE) adducts were identified in mitochondrial ICDH from AAPH-treated cells.

Conclusions:

  • ICDH is vulnerable to oxidative damage from lipid peroxidation products.
  • Damage to ICDH can compromise cellular antioxidant defenses, potentially leading to a prooxidant state.

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