title: Chemical genetic approach identifies microtubule affinity-regulating kinase 1 as a leucine-rich repeat kinase 2 substrate.
abstract: Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of autosomal-dominant forms of Parkinson's disease. LRRK2 is a modular, multidomain protein containing 2 enzymatic domains, including a kinase domain, as well as several protein-protein interaction domains, pointing to a role in cellular signaling. Although enormous efforts have been made, the exact pathophysiologic mechanisms of LRRK2 are still not completely known. In this study, we used a chemical genetics approach to identify LRRK2 substrates from mouse brain. This approach allows the identification of substrates of 1 particular kinase in a complex cellular environment. Several of the identified peptides are involved in the regulation of microtubule (MT) dynamics, including microtubule-associating protein (MAP)/microtubule affinity-regulating kinase 1 (MARK1). MARK1 is a serine/threonine kinase known to phosphorylate MT-binding proteins such as Tau, MAP2, and MAP4 at KXGS motifs leading to MT destabilization. In vitro kinase assays and metabolic-labeling experiments in living cells confirmed MARK1 as an LRRK2 substrate. Moreover, we also showed that LRRK2 and MARK1 are interacting in eukaryotic cells. Our findings contribute to the identification of physiologic LRRK2 substrates and point to a potential mechanism explaining the reported effects of LRRK2 on neurite morphology.
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Petranka
authors: P
authors: Krumova
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Lauran
authors: L
authors: Reyniers
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Marc
authors: M
authors: Meyer
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Evy
authors: E
authors: Lobbestael
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Daniela
authors: D
authors: Stauffer
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Bertran
authors: B
authors: Gerrits
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Lionel
authors: L
authors: Muller
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Sjouke
authors: S
authors: Hoving
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Klemens
authors: K
authors: Kaupmann
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Johannes
authors: J
authors: Voshol
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Doriano
authors: D
authors: Fabbro
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Andreas
authors: A
authors: Bauer
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Giorgio
authors: G
authors: Rovelli
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium jean-marc.taymans@inserm.fr veerle.baekelandt@med.kuleuven.be.
authors: Jean-Marc
authors: JM
authors: Taymans
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium.
authors: Tewis
authors: T
authors: Bouwmeester
authors: *Novartis Institutes for Biomedical Research, Basel, Switzerland; and Department of Neurosciences, Research Group for Neurobiology and Gene Therapy, Katholieke Universiteit (KU) Leuven, Leuven, Belgium jean-marc.taymans@inserm.fr veerle.baekelandt@med.kuleuven.be.
authors: Veerle
authors: V
authors: Baekelandt
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D051379:Mice; D018345:Mice, Knockout; D008870:Microtubules / Q000378:metabolism; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D050505:Mutant Proteins; D011994:Recombinant Proteins; C105547:MARK1 protein, human; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; C495279:Lrrk2 protein, mouse; C531338:MARK1 protein, mouse; D017346:Protein Serine-Threonine Kinases
keywords: Parkinson’s disease; cellular signaling; gatekeeper kinase; neuroscience