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Updated: Sep 20, 2025

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
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Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

Published on: December 14, 2017

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LRRK2 interactions with microtubules are independent of LRRK2-mediated Rab phosphorylation

Tuyana Malankhanova1, Zhiyong Liu1, Enquan Xu1

  • 1Duke Center for Neurodegeneration Research, Department of Pharmacology and Cancer Biology, Duke University, Durham, NC, USA.

EMBO Reports
|May 27, 2025
PubMed

Insights

Microtubules do not directly regulate Parkinson's disease-associated LRRK2 kinase activity. Tubulin and microtubules are transient LRRK2 interactors, not essential for Rab phosphorylation.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Microtubule dysfunction is implicated in neurodegenerative diseases.
  • Parkinson's disease-associated Leucine-rich repeat kinase 2 (LRRK2) is a key protein in neuronal health.
  • Previous studies suggest microtubule interactions may influence LRRK2 kinase activity.

Purpose of the Study:

  • To investigate the direct role of microtubules and tubulin in regulating LRRK2 kinase activity and function.
  • To determine if LRRK2 interacts with microtubules in endogenous systems.

Main Methods:

  • Utilized nocodazole to destabilize microtubules in macrophages with endogenous LRRK2.
  • Assessed LRRK2-mediated Rab phosphorylation following microtubule disruption and GTP supplementation.
  • Employed chemical microtubule stabilization and LRRK2 trapping to membranes.
  • Conducted proximity-labeling proteomics and colocalization studies.
  • Investigated LRRK2 interaction with microtubules using type I inhibitors and N-terminal tags.

Main Results:

  • Nocodazole treatment destabilized microtubules and impaired LRRK2-mediated Rab phosphorylation, which was restored by GTP.
  • Chemical microtubule stabilization had minimal impact on Rab phosphorylation.
  • Trapping LRRK2 to LAMP1-positive membranes upregulated Rab phosphorylation.
  • LRRK2 transiently and independently interacted with both polymerized and free tubulin.
  • Inhibitor-bound LRRK2 did not stably interact with microtubules, unlike tagged LRRK2.

Conclusions:

  • Tubulin and microtubules are transient interacting partners of LRRK2.
  • Microtubule binding is not essential for LRRK2-mediated Rab phosphorylation.
  • Cellular GTP levels, rather than direct microtubule interaction, appear to influence LRRK2 activity.
Keywords:
Cell SignalingCytoskeletonGTPaseParkinson’s DiseaseProtein Kinase

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