title: Alpha-synuclein interacts with Glucocerebrosidase providing a molecular link between Parkinson and Gaucher diseases.
abstract: The presynaptic protein α-synuclein (α-syn), particularly in its amyloid form, is widely recognized for its involvement in Parkinson disease (PD). Recent genetic studies reveal that mutations in the gene GBA are the most widespread genetic risk factor for parkinsonism identified to date. GBA encodes for glucocerebrosidase (GCase), the enzyme deficient in the lysosomal storage disorder, Gaucher disease (GD). In this work, we investigated the possibility of a physical linkage between α-syn and GCase, examining both wild type and the GD-related N370S mutant enzyme. Using fluorescence and nuclear magnetic resonance spectroscopy, we determined that α-syn and GCase interact selectively under lysosomal solution conditions (pH 5.5) and mapped the interaction site to the α-syn C-terminal residues, 118-137. This α-syn-GCase complex does not form at pH 7.4 and is stabilized by electrostatics, with dissociation constants ranging from 1.2 to 22 μm in the presence of 25 to 100 mm NaCl. Intriguingly, the N370S mutant form of GCase has a reduced affinity for α-syn, as does the inhibitor conduritol-β-epoxide-bound enzyme. Immunoprecipitation and immunofluorescence studies verified this interaction in human tissue and neuronal cell culture, respectively. Although our data do not preclude protein-protein interactions in other cellular milieux, we suggest that the α-syn-GCase association is favored in the lysosome, and that this noncovalent interaction provides the groundwork to explore molecular mechanisms linking PD with mutant GBA alleles.
authors.affiliation: Laboratory of Molecular Biophysics, National Heart Lung and Blood Institute, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Thai Leong
authors.initials: TL
authors.lastname: Yap
authors.forename: James M
authors.initials: JM
authors.lastname: Gruschus
authors.forename: Arash
authors.initials: A
authors.lastname: Velayati
authors.forename: Wendy
authors.initials: W
authors.lastname: Westbroek
authors.forename: Ehud
authors.initials: E
authors.lastname: Goldin
authors.forename: Nima
authors.initials: N
authors.lastname: Moaven
authors.forename: Ellen
authors.initials: E
authors.lastname: Sidransky
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Lee
mesh_terms: D019943:Amino Acid Substitution; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005776:Gaucher Disease / Q000235:genetics / Q000378:metabolism*; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007294:Inositol / Q000031:analogs & derivatives / Q000494:pharmacology; D008247:Lysosomes / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D020125:Mutation, Missense; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism*; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D046912:Multiprotein Complexes; D051844:alpha-Synuclein; D007294:Inositol; D005962:Glucosylceramidase; C011423:conduritol epoxide