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Related Experiment Video

Updated: Apr 13, 2026

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
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LRRK2 dephosphorylation increases its ubiquitination.

Jing Zhao1, Tyler P Molitor1, J William Langston1

  • 1The Parkinson's Institute, 675 Almanor Ave, Sunnyvale, CA 94085, U.S.A.

The Biochemical Journal
|May 6, 2015
PubMed
Summary

Inhibiting leucine rich repeat protein kinase 2 (LRRK2) causes dephosphorylation and subsequent ubiquitination and degradation of LRRK2. This dephosphorylation-ubiquitination cycle may explain loss-of-function Parkinson's disease phenotypes.

Keywords:
Parkinson's diseasekinasekinase inhibitorphosphataseubiquitin

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Activating mutations in the leucine rich repeat protein kinase 2 (LRRK2) gene are a common cause of inherited Parkinson's disease (PD).
  • LRRK2 phosphorylation sites, including Ser935, are dephosphorylated in PD mutants and upon kinase inhibition, leading to loss of 14-3-3 binding.
  • The consequences of LRRK2 dephosphorylation are not fully understood.

Purpose of the Study:

  • To investigate the consequences of LRRK2 dephosphorylation induced by kinase inhibition.
  • To explore the link between LRRK2 dephosphorylation and ubiquitination.
  • To elucidate the role of phosphatase activity in LRRK2 ubiquitination.

Main Methods:

  • Treatment with a potent and selective LRRK2 inhibitor (GNE1023).
  • Analysis of LRRK2 phosphorylation, ubiquitination, and stability in expression systems, cell lines (A549), and mouse models.
  • Investigation of LRRK2 ubiquitination in PD mutants and in cells with blocked 14-3-3 binding (difopein expression).
  • Assessment of the effect of Calyculin A on LRRK2 dephosphorylation and ubiquitination.

Main Results:

  • LRRK2 kinase inhibition led to Ser935 dephosphorylation, followed by LRRK2 ubiquitination and degradation.
  • GNE1023 treatment decreased LRRK2 phosphorylation and stability in various systems.
  • LRRK2 ubiquitination involved Lys48 and Lys63 linkages.
  • PD-associated LRRK2 mutants and blocked 14-3-3 binding resulted in LRRK2 hyper-ubiquitination.
  • Calyculin A prevented dephosphorylation and reduced ubiquitination, implicating phosphatase activity.

Conclusions:

  • LRRK2 inhibition triggers a dephosphorylation-ubiquitination cascade leading to LRRK2 degradation.
  • This dynamic cycle is linked to phosphatase activity and may explain loss-of-function phenotypes in PD.
  • Understanding this cycle offers insights into Parkinson's disease pathogenesis and potential therapeutic strategies.