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Cited by 13Protein Kinase Regulation and GTPase SignalingParkinson's Disease Mechanisms and TreatmentsProtein Tyrosine Phosphatases

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PKC isoforms activate LRRK1 kinase by phosphorylating conserved residues (Ser1064, Ser1074 and Thr1075) within the CORB GTPase domain

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Malik, Asad U., Karapetsas, Athanasios, Nirujogi, Raja S., Chatterjee, Deep, Phung, Toan K., Wightman, Melanie, Gourlay, Robert, Morrice, Nick, Mathea, Sebastian, Knapp, Stefan, Alessi, Dario R.·2022·Biochemical Journal
DOI 10.1042/bcj20220308PMC PMC9555798

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Abstract

Leucine-rich-repeat-kinase 1 (LRRK1) and its homolog LRRK2 are multidomain kinases possessing a ROC-CORA-CORB containing GTPase domain and phosphorylate distinct Rab proteins. LRRK1 loss of function mutations cause the bone disorder osteosclerotic metaphyseal dysplasia, whereas LRRK2 missense mutations that enhance kinase activity cause Parkinson's disease. Previous work suggested that LRRK1 but not LRRK2, is activated via a Protein Kinase C (PKC)-dependent mechanism. Here we demonstrate that phosphorylation and activation of LRRK1 in HEK293 cells is blocked by PKC inhibitors including LXS-196 (Darovasertib), a compound that has entered clinical trials. We show multiple PKC isoforms phosphorylate and activate recombinant LRRK1 in a manner reversed by phosphatase treatment. PKCα unexpectedly does not activate LRRK1 by phosphorylating the kinase domain, but instead phosphorylates a cluster of conserved residues (Ser1064, Ser1074 and Thr1075) located within a region of the CORB domain of the GTPase domain. These residues are positioned at the equivalent region of the LRRK2 DK helix reported to stabilize the kinase domain αC-helix in the active conformation. Thr1075 represents an optimal PKC site phosphorylation motif and its mutation to Ala, blocked PKC-mediated activation of LRRK1. A triple Glu mutation of Ser1064/Ser1074/Thr1075 to mimic phosphorylation, enhanced LRRK1 kinase activity ∼3-fold. From analysis of available structures, we postulate that phosphorylation of Ser1064, Ser1074 and Thr1075 activates LRRK1 by promoting interaction and stabilization of the αC-helix on the kinase domain. This study provides new fundamental insights into the mechanism controlling LRRK1 activity and reveals a novel unexpected activation mechanism.

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Metadata

Authors
Malik, A. U., Karapetsas, A., Nirujogi, R. S., Chatterjee, D., Phung, T. K., Wightman, M., Gourlay, R., Morrice, N., Mathea, S., Knapp, S., & Alessi, D. R.
Affiliations
  • University of Dundee (GB)
  • Goethe University Frankfurt (DE)
  • 4AB Sciex, Alderley Park, Macclesfield SK10 4TG, U.K.
Corresponding author
Alessi, Dario R.
ORCID
Venue
Biochemical Journal
Publisher
Portland Press Ltd.
Published
2022-09-23
Volume
479
Issue
18
Pages
1941-1965
ISSN
  • 0264-6021 (ISSN-L)
  • 1470-8728
DOI
10.1042/bcj20220308
PMC
PMC9555798
OpenAlex
W4293567400
Cited by
13
References
1
Citations by year
2026: 3 · 2025: 5 · 2024: 1 · 2023: 3 · 2022: 1
FWCI
0.82
Type
journal article
Language
en
Open access
Licence
Locations
  • Biochemical Journal (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Discovery Research Portal (University of Dundee) (submittedVersion, cc-by)
Funders
  • Michael J. Fox Foundation for Parkinson's Research (ASAP‐000463)
  • GlaxoSmithKline
  • Merck KGaA
  • University of California, San Diego
  • Medical Research Council (MC_UU_00018/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Health Sciences › Medicine › Clinical Neurology › Parkinson's Disease Mechanisms and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
Keywords
LRRK1, LRRK2, protein kinase C, kinase activation, GTPase domain, phosphorylation, αC-helix, Parkinson's disease
MeSH terms
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2, Humans, Leucine, Mutation, Phosphoric Monoester Hydrolases, Phosphorylation, Protein Kinase C, Protein Serine-Threonine Kinases, Protein Isoforms, GTP Phosphohydrolases, Cordyceps, Protein Kinase Inhibitors, Protein Kinase C-alpha, HEK293 Cells

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