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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
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A mitochondria-targeted derivative of ascorbate: MitoC.

Peter G Finichiu1, David S Larsen2, Cameron Evans2

  • 1MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.

Free Radical Biology & Medicine
|October 11, 2015
PubMed
Summary

Researchers developed MitoC, a targeted antioxidant for mitochondria. This novel approach effectively combats mitochondrial oxidative damage, offering new therapeutic potential for related diseases.

Keywords:
Ascorbic acidLipid peroxidationLipophilic cationMitoCMitoPeroxMitochondriaMitochondrial targeting

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Pharmacology

Background:

  • Mitochondrial oxidative damage is implicated in numerous pathologies.
  • Targeting antioxidants to mitochondria via triphenylphosphonium (TPP) cation is a therapeutic strategy.
  • Current methods primarily target hydrophobic antioxidants, limiting options for hydrophilic compounds.

Purpose of the Study:

  • To develop and characterize MitoC, a mitochondria-targeted version of the hydrophilic antioxidant ascorbate.
  • To investigate the feasibility of targeting polar and acidic compounds to mitochondria.
  • To evaluate MitoC's efficacy as a mitochondrial antioxidant.

Main Methods:

  • Conjugation of ascorbate to the triphenylphosphonium (TPP) cation via a hydrophobic linker.
  • Assessment of MitoC uptake and localization within mitochondria.
  • Evaluation of MitoC's antioxidant activity against reactive species and lipid peroxidation.
  • Analysis of MitoC's recycling within mitochondria via cellular redox systems.

Main Results:

  • MitoC successfully targets and accumulates within mitochondria, overcoming ascorbate's polarity and acidity.
  • MitoC effectively scavenges reactive oxygen species and prevents mitochondrial lipid peroxidation.
  • MitoC is rapidly recycled to active ascorbate by mitochondrial glutathione and thioredoxin systems.
  • MitoC protects aconitase from superoxide-induced inactivation.

Conclusions:

  • The TPP moiety can effectively target polar and acidic compounds, like ascorbate, to mitochondria.
  • MitoC demonstrates significant therapeutic potential as a novel mitochondria-targeted antioxidant.
  • MitoC serves as a valuable research tool for studying mitochondrial ascorbate function.