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Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
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PAK6-mediated phosphorylation of PPP2R2C regulates LRRK2-PP2A complex formation

Lucia Iannotta1,2, Marco Emanuele3, Giulia Favetta1

  • 1Department of Biology, University of Padova, Padua, Italy.

Frontiers in Molecular Neuroscience
|January 3, 2024
PubMed

Insights

Leucine-rich repeat kinase 2 (LRRK2) phosphorylation is linked to Parkinson's disease. PAK6 kinase phosphorylates PPP2R2C, influencing its LRRK2 binding and localization, revealing a new regulatory mechanism for LRRK2 activity.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Neuroscience

Background:

  • Mutations in leucine-rich repeat kinase 2 (LRRK2) are a significant cause of Parkinson's disease (PD).
  • Dephosphorylation of LRRK2 at specific sites is implicated in PD pathogenesis.
  • Previous studies identified PP1, PP2A phosphatases, and PAK6 kinase as LRRK2 regulators.

Purpose of the Study:

  • To investigate a potential functional link between PAK6 kinase and LRRK2 phosphatases.
  • To elucidate the role of PAK6-mediated phosphorylation of the PP2A regulatory subunit PPP2R2C in LRRK2 regulation.

Main Methods:

  • PhosTag gel electrophoresis was employed using purified proteins.
  • Investigated the effect of PPP2R2C phosphorylation at S381 on PP2A holoenzyme formation and LRRK2 binding.
  • Assessed the impact of PAK6 kinase activity on PPP2R2C subcellular localization.
  • Evaluated the effect of PPP2R2C phosphorylation on PAK6-mediated LRRK2 dephosphorylation.

Main Results:

  • PAK6 phosphorylates PPP2R2C at serine 381 (S381).
  • S381 phosphorylation of PPP2R2C impairs its binding to LRRK2 but does not affect PP2A holoenzyme formation.
  • PAK6 kinase activity alters PPP2R2C subcellular localization in a manner dependent on S381 phosphorylation.
  • PAK6-mediated LRRK2 dephosphorylation is independent of PPP2R2C S381 phosphorylation, suggesting a dominant 14-3-3 mediated mechanism.

Conclusions:

  • PAK6-mediated phosphorylation of PPP2R2C influences its recruitment to LRRK2 and subcellular localization.
  • This finding reveals an additional layer of regulation in the fine-tuning of LRRK2 phosphorylation.
  • The study provides insights into the complex molecular mechanisms underlying Parkinson's disease.
Keywords:
LRRK2PP2APPP2R2CPak6Parkinson’s diseasedephosphorylationphosphorylation

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