title: Structural Characterization of LRRK2 Inhibitors.
abstract: Kinase inhibition is considered to be an important therapeutic target for LRRK2 mediated Parkinson's disease (PD). Many LRRK2 kinase inhibitors have been reported but have yet to be optimized in order to qualify as drug candidates for the treatment of the disease. In order to start a structure-function analysis of such inhibitors, we mutated the active site of Dictyostelium Roco4 kinase to resemble LRRK2. Here, we show saturation transfer difference (STD) NMR and the first cocrystal structures of two potent in vitro inhibitors, LRRK2-IN-1 and compound 19, with mutated Roco4. Our data demonstrate that this system can serve as an excellent tool for the structural characterization and optimization of LRRK2 inhibitors using X-ray crystallography and NMR spectroscopy.
authors.affiliation: †Department of Cell Biochemistry, University of Groningen, 9747AG Groningen, The Netherlands.
authors.forename: Bernd K
authors.initials: BK
authors.lastname: Gilsbach
authors.affiliation: ‡Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.|§Center for Integrated Protein Science Munich at Biomolecular NMR Spectroscopy, Department Chemie, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching bei München, Germany.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Messias
authors.affiliation: ∥University of Dundee, DD1 4HN Dundee, Scotland.
authors.forename: Genta
authors.initials: G
authors.lastname: Ito
authors.affiliation: ‡Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.|§Center for Integrated Protein Science Munich at Biomolecular NMR Spectroscopy, Department Chemie, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching bei München, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Sattler
authors.affiliation: ∥University of Dundee, DD1 4HN Dundee, Scotland.
authors.forename: Dario R
authors.initials: DR
authors.lastname: Alessi
authors.affiliation: ⊥Max-Planck Institut für Molekulare Physiologie, 44202 Dortmund, Germany.
authors.forename: Alfred
authors.initials: A
authors.lastname: Wittinghofer
authors.affiliation: †Department of Cell Biochemistry, University of Groningen, 9747AG Groningen, The Netherlands.
authors.forename: Arjan
authors.initials: A
authors.lastname: Kortholt
mesh_terms: D018360:Crystallography, X-Ray; D004023:Dictyostelium / Q000201:enzymology*; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D062105:Molecular Docking Simulation; D009025:Morpholines / Q000737:chemistry; D017354:Point Mutation; D011485:Protein Binding; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D011743:Pyrimidines / Q000737:chemistry
chemical_list: D009025:Morpholines; D015800:Protozoan Proteins; D011743:Pyrimidines; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases