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Updated: Sep 20, 2025
Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
Published on: December 14, 2017
Distinct profiles of LRRK2 activation and Rab GTPase phosphorylation in clinical samples from different PD cohorts
Lilian Petropoulou-Vathi1, Athina Simitsi2, Politymi-Eleni Valkimadi1
1Center for Clinical, Experimental Surgery, and Translational Research, Biomedical Research Foundation of the Academy of Athens, Athens, Greece.
Abstract:
Despite several advances in the field, pharmacodynamic outcome measures reflective of LRRK2 kinase activity in clinical biofluids remain urgently needed. A variety of targets and approaches have been utilized including assessments of LRRK2 itself (levels, phosphorylation), or its substrates (e.g. Rab10 or other Rab GTPases). We have previously shown that intrinsic kinase activity of LRRK2 isolated from PBMCs of G2019S carriers is elevated, irrespective of disease status. In the present study we find that phosphorylation of Rab10 is also elevated in G2019S carriers, but only those with PD. Additionally, phosphorylation of this substrate is also elevated in two separate idiopathic PD cohorts, but not in carriers of the A53T mutation in α-synuclein. In contrast, Rab29 phosphorylation was specifically reduced in urinary exosomes from A53T and idiopathic PD patients. Taken together, our findings highlight the need for the assessment of multiple complimentary targets for a more comprehensive picture of the disease.
Insights
Measuring LRRK2 kinase activity in Parkinson's disease (PD) is crucial. This study found Rab10 phosphorylation elevated in G2019S and idiopathic PD, while Rab29 phosphorylation decreased in A53T and idiopathic PD urinary exosomes.
Area of Science:
- Neuroscience
- Genetics
- Biochemistry
Background:
- Pharmacodynamic outcome measures for LRRK2 kinase activity in biofluids are needed for Parkinson's disease (PD) research.
- Previous work showed elevated LRRK2 kinase activity in G2019S carriers, irrespective of disease status.
Purpose of the Study:
- To investigate LRRK2 substrate phosphorylation in clinical biofluids as potential PD biomarkers.
- To assess Rab10 and Rab29 phosphorylation in relation to LRRK2 mutations (G2019S, A53T) and idiopathic PD.
Main Methods:
- Analysis of Rab10 and Rab29 phosphorylation levels in peripheral blood mononuclear cells (PBMCs) and urinary exosomes.
- Comparison of phosphorylation patterns across different patient cohorts: G2019S carriers (with and without PD), idiopathic PD patients, and A53T carriers.
Main Results:
- Rab10 phosphorylation was elevated in G2019S carriers with PD and in idiopathic PD cohorts.
- Rab10 phosphorylation was not elevated in A53T mutation carriers.
- Rab29 phosphorylation was reduced in urinary exosomes from A53T and idiopathic PD patients.
Conclusions:
- Rab10 phosphorylation serves as a potential pharmacodynamic marker for LRRK2 activity in G2019S and idiopathic PD.
- Rab29 phosphorylation changes in urinary exosomes may indicate distinct pathological mechanisms in A53T and idiopathic PD.
- Assessing multiple targets, including Rab10 and Rab29, provides a more comprehensive understanding of LRRK2 pathway involvement in PD.
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