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Hydrogen peroxide-dependent oxidation of ERK2 within its D-recruitment site alters its substrate selection

Anthony E Postiglione1,2, Laquaundra L Adams1, Ese S Ekhator1

  • 1Department of Biology, North Carolina A&T State University, Greensboro, NC 27411, USA.

Iscience
|September 25, 2023
PubMed

Insights

Oxidation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) by hydrogen peroxide alters substrate interactions. This redox modification impacts ERK1/2

Area of Science:

  • Biochemistry
  • Cell Biology
  • Redox Biology

Background:

  • Extracellular signal-regulated kinases 1 and 2 (ERK1/2) are crucial signaling proteins implicated in numerous diseases.
  • ERK1/2 activity is regulated by phosphorylation, but recent findings indicate redox modifications also play a role.
  • Signal-generated hydrogen peroxide has been identified as an oxidant that can modify ERK1/2.

Purpose of the Study:

  • To investigate how oxidation of ERK2 by hydrogen peroxide affects its interaction with model substrates.
  • To elucidate the mechanism by which redox modification influences ERK2's substrate selectivity.
  • To explore the functional consequences of ERK1/2 oxidation on kinase activity in cellular contexts.

Main Methods:

  • Biochemical assays to assess enzyme activity and substrate binding.
  • Computational modeling to predict and understand structural changes upon oxidation.
  • Site-directed mutagenesis to identify critical residues involved in oxidation and substrate interaction.
  • Cell-based assays using HeLa cells to examine ERK1/2 activity in vivo.

Main Results:

  • ERK2 undergoes sulfenylation at cysteine 159 (C159) on its D-recruitment site.
  • Oxidation at C159 differentially modulates ERK2's affinity for various substrates, decreasing it for some and increasing it for others.
  • Peroxide-dependent oxidation provides a mechanism for altered ERK2-substrate interactions.
  • Oxidized ERK1/2 exhibits enhanced phosphorylation of ribosomal S6 kinase A1 (RSK1) in HeLa cells.

Conclusions:

  • Redox modification of ERK1/2, specifically sulfenylation at C159, directly impacts its interaction with substrates.
  • Oxidation alters ERK2's substrate binding affinity in a substrate-dependent manner.
  • These findings reveal a crosstalk mechanism between redox and phosphorylation signaling pathways at the level of kinase-substrate selection.
  • This study provides a foundation for understanding how redox state influences kinase signaling specificity.
Keywords:
BiochemistryBiological sciencesCell biologyMolecular biology

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