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ROS-dependent localization of glycolytic enzymes to mitochondria.

Pau B Esparza-Moltó1, Arvind V Goswami1, Süleyman Bozkurt2

  • 1Salk Institute for Biological Studies, La Jolla, CA, 92037, USA.

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Mitochondrial reactive oxygen species (mtROS) signaling involves dynamic protein changes. This study found glycolytic enzymes increase mitochondrial localization under oxidative stress, suggesting an adaptive response.

Keywords:
Alzheimer's diseaseGlycolytic enzymesMitochondriaProximity labelingReactive oxygen speciesStress signaling

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Area of Science:

  • Mitochondrial biology
  • Cellular signaling
  • Oxidative stress

Background:

  • Mitochondrial reactive oxygen species (mtROS) play dual roles in cellular signaling and oxidative stress.
  • Dysregulated mtROS are implicated in aging and neurodegenerative diseases.
  • Protein redistribution between cellular compartments is a key regulatory mechanism.

Purpose of the Study:

  • To identify proteins with dynamic mitochondrial localization in response to increased mtROS production.
  • To investigate the role of glycolytic enzymes in mitochondrial oxidative stress signaling.
  • To explore the adaptive significance of altered glycolytic enzyme localization.

Main Methods:

  • Used BirA* biotin ligase targeted to the outer mitochondrial membrane in HEK293 cells for proximity-dependent labeling.
  • Treated cells with menadione to induce mtROS production.
  • Employed mitochondrial fractionation, imaging, and submitochondrial fractionation to confirm protein localization.
  • Analyzed primary Alzheimer's disease fibroblasts.

Main Results:

  • Identified proteins with altered mitochondrial localization upon menadione treatment.
  • Confirmed increased mitochondrial localization of glycolytic enzymes in response to menadione, hypoxia, and a mitochondria-targeted oxidase, independent of total protein levels.
  • Observed inhibition of this localization by the antioxidant MnTBAP.
  • Found reduced mitochondrial association of glycolytic enzymes in Alzheimer's disease fibroblasts, responsive to antioxidants.

Conclusions:

  • Increased mitochondrial localization of glycolytic enzymes is an adaptive response to mtROS.
  • This redistribution may alter glucose metabolism, create new metabolic circuits, or confer cytoprotection.
  • Further research into these enzymes' moonlighting functions and roles in oxidative stress is warranted.