title: Characterization of the complex formed by β-glucocerebrosidase and the lysosomal integral membrane protein type-2.
abstract: The lysosomal integral membrane protein type-2 (LIMP-2) plays a pivotal role in the delivery of β-glucocerebrosidase (GC) to lysosomes. Mutations in GC result in Gaucher's disease (GD) and are the major genetic risk factor for the development of Parkinson's disease (PD). Variants in the LIMP-2 gene cause action myoclonus-renal failure syndrome and also have been linked to PD. Given the importance of GC and LIMP-2 in disease pathogenesis, we studied their interaction sites in more detail. Our previous data demonstrated that the crystal structure of LIMP-2 displays a hydrophobic three-helix bundle composed of helices 4, 5, and 7, of which helix 5 and 7 are important for ligand binding. Here, we identified a similar helical motif in GC through surface potential analysis. Coimmunoprecipitation and immunofluorescence studies revealed a triple-helical interface region within GC as critical for LIMP-2 binding and lysosomal transport. Based on these findings, we generated a LIMP-2 helix 5-derived peptide that precipitated and activated recombinant wild-type and GD-associated N370S mutant GC in vitro. The helix 5 peptide fused to a cell-penetrating peptide also activated endogenous lysosomal GC and reduced α-synuclein levels, suggesting that LIMP-2-derived peptides can be used to activate endogenous as well as recombinant wild-type or mutant GC efficiently. Our data also provide a structural model of the LIMP-2/GC complex that will facilitate the development of GC chaperones and activators as potential therapeutics for GD, PD, and related synucleinopathies.
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Friederike
authors.initials: F
authors.lastname: Zunke
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Lisa
authors.initials: L
authors.lastname: Andresen
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Sophia
authors.initials: S
authors.lastname: Wesseler
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Johann
authors.initials: J
authors.lastname: Groth
authors.affiliation: Anatomical Institute, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Philipp
authors.initials: P
authors.lastname: Arnold
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Michelle
authors.initials: M
authors.lastname: Rothaug
authors.affiliation: Department of Neurology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611;
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Mazzulli
authors.affiliation: Department of Neurology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611;
authors.forename: Dimitri
authors.initials: D
authors.lastname: Krainc
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Judith
authors.initials: J
authors.lastname: Blanz
authors.affiliation: Institute of Biochemistry, Christian-Albrechts-University of Kiel, 24098 Kiel, Germany;
authors.forename: Paul
authors.initials: P
authors.lastname: Saftig
authors.affiliation: Faculty of Chemistry/Biochemistry III, University Bielefeld, 33615 Bielefeld, Germany michael.schwake@uni-bielefeld.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Schwake
mesh_terms: D020816:Amino Acid Motifs / Q000502:physiology; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D005962:Glucosylceramidase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D052119:Lysosomal-Associated Membrane Protein 2 / Q000235:genetics / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D017434:Protein Structure, Tertiary
chemical_list: D052119:Lysosomal-Associated Membrane Protein 2; D005962:Glucosylceramidase
keywords: GC activators; Gaucher's disease; LIMP-2; Parkinson's disease; β-glucocerebrosidase