title: Structural model of the dimeric Parkinson's protein LRRK2 reveals a compact architecture involving distant interdomain contacts.
abstract: Leucine-rich repeat kinase 2 (LRRK2) is a large, multidomain protein containing two catalytic domains: a Ras of complex proteins (Roc) G-domain and a kinase domain. Mutations associated with familial and sporadic Parkinson's disease (PD) have been identified in both catalytic domains, as well as in several of its multiple putative regulatory domains. Several of these mutations have been linked to increased kinase activity. Despite the role of LRRK2 in the pathogenesis of PD, little is known about its overall architecture and how PD-linked mutations alter its function and enzymatic activities. Here, we have modeled the 3D structure of dimeric, full-length LRRK2 by combining domain-based homology models with multiple experimental constraints provided by chemical cross-linking combined with mass spectrometry, negative-stain EM, and small-angle X-ray scattering. Our model reveals dimeric LRRK2 has a compact overall architecture with a tight, multidomain organization. Close contacts between the N-terminal ankyrin and C-terminal WD40 domains, and their proximity-together with the LRR domain-to the kinase domain suggest an intramolecular mechanism for LRRK2 kinase activity regulation. Overall, our studies provide, to our knowledge, the first structural framework for understanding the role of the different domains of full-length LRRK2 in the pathogenesis of PD.
authors.affiliation: German Center for Neurodegenerative Diseases, 72076 Tübingen, Germany; Center for Ophthalmology, Institute for Ophthalmic Research, Eberhard Karls University, 72076 Tübingen, Germany;
authors.forename: Giambattista
authors.initials: G
authors.lastname: Guaitoli
authors.affiliation: Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy; Cell Networks, University of Heidelberg, 69120 Heidelberg, Germany;
authors.forename: Francesco
authors.initials: F
authors.lastname: Raimondi
authors.affiliation: German Center for Neurodegenerative Diseases, 72076 Tübingen, Germany; Department of Cell Biochemistry, University of Groningen, Groningen 9747 AG, The Netherlands; Structural Biology Group, Max Planck Institute for Molecular Physiology, 44227 Dortmund, Germany;
authors.forename: Bernd K
authors.initials: BK
authors.lastname: Gilsbach
authors.affiliation: Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029;
authors.forename: Yacob
authors.initials: Y
authors.lastname: Gómez-Llorente
authors.affiliation: Structural Biology Brussels, Vrije Universiteit Brussel, 1050 Brussels, Belgium; Vlaams Instituut voor Biotechnologie, Structural Biology Research Center, Vrije Universiteit Brussel, 1050 Brussels, Belgium;
authors.forename: Egon
authors.initials: E
authors.lastname: Deyaert
authors.affiliation: Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029;
authors.forename: Fabiana
authors.initials: F
authors.lastname: Renzi
authors.affiliation: Departments of Neurology and Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029;
authors.forename: Xianting
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029; Departments of Neurology and Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029;
authors.forename: Adam
authors.initials: A
authors.lastname: Schaffner
authors.affiliation: Center for Integrated Protein Science Munich at Department of Chemistry, Technische Universität München, 85747 Garching, Germany; Institute of Structural Biology, Helmholtz Zentrum München, 85764 Munich, Germany;
authors.forename: Pravin Kumar Ankush
authors.initials: PK
authors.lastname: Jagtap
authors.affiliation: Center for Ophthalmology, Institute for Ophthalmic Research, Eberhard Karls University, 72076 Tübingen, Germany;
authors.forename: Karsten
authors.initials: K
authors.lastname: Boldt
authors.affiliation: German Center for Neurodegenerative Diseases, 72076 Tübingen, Germany; Center for Ophthalmology, Institute for Ophthalmic Research, Eberhard Karls University, 72076 Tübingen, Germany;
authors.forename: Felix
authors.initials: F
authors.lastname: von Zweydorf
authors.affiliation: Structural Biology Group, Max Planck Institute for Molecular Physiology, 44227 Dortmund, Germany;
authors.forename: Katja
authors.initials: K
authors.lastname: Gotthardt
authors.affiliation: Beryllium Discovery Corporation, Bainbridge Island, WA 98110;
authors.forename: Donald D
authors.initials: DD
authors.lastname: Lorimer
authors.affiliation: Departments of Neurology and Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029;
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Yue
authors.affiliation: Broad Institute, Cambridge, MA 02142;
authors.forename: Alex
authors.initials: A
authors.lastname: Burgin
authors.affiliation: SomaLogic, Boulder, CO 80301.
authors.forename: Nebojsa
authors.initials: N
authors.lastname: Janjic
authors.affiliation: Center for Integrated Protein Science Munich at Department of Chemistry, Technische Universität München, 85747 Garching, Germany; Institute of Structural Biology, Helmholtz Zentrum München, 85764 Munich, Germany;
authors.forename: Michael
authors.initials: M
authors.lastname: Sattler
authors.affiliation: Structural Biology Brussels, Vrije Universiteit Brussel, 1050 Brussels, Belgium; Vlaams Instituut voor Biotechnologie, Structural Biology Research Center, Vrije Universiteit Brussel, 1050 Brussels, Belgium;
authors.forename: Wim
authors.initials: W
authors.lastname: Versées
authors.affiliation: Center for Ophthalmology, Institute for Ophthalmic Research, Eberhard Karls University, 72076 Tübingen, Germany;
authors.forename: Marius
authors.initials: M
authors.lastname: Ueffing
authors.affiliation: Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029; iban.ubarretxena@mssm.edu a.kortholt@rug.nl johannes.gloeckner@dzne.de.
authors.forename: Iban
authors.initials: I
authors.lastname: Ubarretxena-Belandia
authors.affiliation: Department of Cell Biochemistry, University of Groningen, Groningen 9747 AG, The Netherlands; iban.ubarretxena@mssm.edu a.kortholt@rug.nl johannes.gloeckner@dzne.de.
authors.forename: Arjan
authors.initials: A
authors.lastname: Kortholt
authors.affiliation: German Center for Neurodegenerative Diseases, 72076 Tübingen, Germany; Center for Ophthalmology, Institute for Ophthalmic Research, Eberhard Karls University, 72076 Tübingen, Germany; iban.ubarretxena@mssm.edu a.kortholt@rug.nl johannes.gloeckner@dzne.de.
authors.forename: Christian Johannes
authors.initials: CJ
authors.lastname: Gloeckner
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D057809:HEK293 Cells; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism; D000072417:Protein Domains; D055503:Protein Multimerization; D017386:Sequence Homology, Amino Acid
chemical_list: D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
keywords: CL-MS; EM; LRRK2; Parkinson’s disease; structural modeling