title: Phosphoproteomics reveals that Parkinson's disease kinase LRRK2 regulates a subset of Rab GTPases.
abstract: Mutations in Park8, encoding for the multidomain Leucine-rich repeat kinase 2 (LRRK2) protein, comprise the predominant genetic cause of Parkinson's disease (PD). G2019S, the most common amino acid substitution activates the kinase two- to threefold. This has motivated the development of LRRK2 kinase inhibitors; however, poor consensus on physiological LRRK2 substrates has hampered clinical development of such therapeutics. We employ a combination of phosphoproteomics, genetics, and pharmacology to unambiguously identify a subset of Rab GTPases as key LRRK2 substrates. LRRK2 directly phosphorylates these both in vivo and in vitro on an evolutionary conserved residue in the switch II domain. Pathogenic LRRK2 variants mapping to different functional domains increase phosphorylation of Rabs and this strongly decreases their affinity to regulatory proteins including Rab GDP dissociation inhibitors (GDIs). Our findings uncover a key class of bona-fide LRRK2 substrates and a novel regulatory mechanism of Rabs that connects them to PD.
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Steger
authors.affiliation: Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors.forename: Francesca
authors.initials: F
authors.lastname: Tonelli
authors.affiliation: Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors.forename: Genta
authors.initials: G
authors.lastname: Ito
authors.affiliation: Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Davies
authors.affiliation: Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors.forename: Matthias
authors.initials: M
authors.lastname: Trost
authors.affiliation: Department of Structural Cell Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Vetter
authors.affiliation: Department of Structural Cell Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Wachter
authors.affiliation: Department of Structural Cell Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Esben
authors.initials: E
authors.lastname: Lorentzen
authors.affiliation: Molecular Discovery Research, GlaxoSmithKline Pharmaceuticals R&D, Harlow, United Kingdom.
authors.forename: Graham
authors.initials: G
authors.lastname: Duddy
authors.affiliation: RD Platform Technology and Science, GlaxoSmithKline Pharmaceuticals R&D, Stevenage, United Kingdom.
authors.forename: Stephen
authors.initials: S
authors.lastname: Wilson
authors.affiliation: The Michael J. Fox Foundation for Parkinson's Research, New York, United States.
authors.forename: Marco As
authors.initials: MA
authors.lastname: Baptista
authors.affiliation: The Michael J. Fox Foundation for Parkinson's Research, New York, United States.
authors.forename: Brian K
authors.initials: BK
authors.lastname: Fiske
authors.affiliation: Early Discovery Neuroscience, Merck Research Laboratories, Boston, United States.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Fell
authors.affiliation: Neuroscience, Merck Research Laboratories, Westpoint, United States.
authors.forename: John A
authors.initials: JA
authors.lastname: Morrow
authors.affiliation: Neurodegeneration Discovery Performance Unit, GlaxoSmithKline Pharmaceuticals R&D, Stevenage, United Kingdom.
authors.forename: Alastair D
authors.initials: AD
authors.lastname: Reith
authors.affiliation: Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors.forename: Dario R
authors.initials: DR
authors.lastname: Alessi
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
mesh_terms: D000818:Animals; D005786:Gene Expression Regulation; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / Q000378:metabolism*; D018345:Mice, Knockout; D010300:Parkinson Disease / Q000503:physiopathology*; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis*; D020691:rab GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D020543:Proteome; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D020691:rab GTP-Binding Proteins
keywords: LRRK2; biochemistry; cell biology; chemical proteomics; human; mouse; neurodegeneration; parkinson-s disease; phosphoproteomics; rabs
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