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PMID: 21653695 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

Alpha-synuclein interacts with Glucocerebrosidase providing a molecular link between Parkinson and Gaucher diseases.

The Journal of biological chemistry ·Vol. 286 ·No. 32 ·2011-08-12 ·Pages 28080-8

Yap TL, Gruschus JM, Velayati A, Westbroek W, Goldin E, Moaven N, Sidransky E, Lee JC

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.

MeSH Terms
Amino Acid Substitution Cell Line, Tumor Enzyme Inhibitors/pharmacology Gaucher Disease/genetics,metabolism Glucosylceramidase/antagonists & inhibitors,genetics,metabolism Humans Hydrogen-Ion Concentration Inositol/analogs & derivatives,pharmacology Lysosomes/genetics,metabolism Multiprotein Complexes/genetics,metabolism Mutation, Missense Parkinson Disease/genetics,metabolism alpha-Synuclein/genetics,metabolism
Chemicals
Enzyme Inhibitors Multiprotein Complexes alpha-Synuclein Inositol Glucosylceramidase conduritol epoxide
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Yap Thai Leong
Laboratory of Molecular Biophysics, National Heart Lung and Blood Institute, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Gruschus James M
Velayati Arash
Westbroek Wendy
Goldin Ehud
Moaven Nima
Sidransky Ellen
Lee Jennifer C
References (75)
75 references, click to expand
  1. Structure of membrane-bound alpha-synuclein from site-directed spin labeling and computational refinement.
    Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):19666-71 PMID: 19066219
  2. The role of autophagy-lysosome pathway in neurodegeneration associated with Parkinson's disease.
    Brain. 2008 Aug;131(Pt 8):1969-78 PMID: 18187492
  3. Impaired degradation of mutant alpha-synuclein by chaperone-mediated autophagy.
    Science. 2004 Aug 27;305(5688):1292-5 PMID: 15333840
  4. Mutation in the alpha-synuclein gene identified in families with Parkinson's disease.
    Science. 1997 Jun 27;276(5321):2045-7 PMID: 9197268
  5. alpha-Synuclein and neuronal cell death.
    Mol Neurodegener. 2009 Feb 04;4:9 PMID: 19193223
  6. Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism.
    Science. 2003 Jan 10;299(5604):256-9 PMID: 12446870
  7. GM1 specifically interacts with alpha-synuclein and inhibits fibrillation.
    Biochemistry. 2007 Feb 20;46(7):1868-77 PMID: 17253773
  8. alpha-Synuclein locus triplication causes Parkinson's disease.
    Science. 2003 Oct 31;302(5646):841 PMID: 14593171
  9. Glucocerebrosidase mutations are an important risk factor for Lewy body disorders.
    Neurology. 2006 Sep 12;67(5):908-10 PMID: 16790605
  10. On the mechanism of internalization of alpha-synuclein into microglia: roles of ganglioside GM1 and lipid raft.
    J Neurochem. 2009 Jul;110(1):400-11 PMID: 19457104
  11. The association of alpha-synuclein with membranes affects bilayer structure, stability, and fibril formation.
    J Biol Chem. 2003 Oct 10;278(41):40186-97 PMID: 12885775
  12. Phosphorylation, lipid raft interaction and traffic of alpha-synuclein in a yeast model for Parkinson.
    Biochim Biophys Acta. 2008 Oct;1783(10):1767-80 PMID: 18634833
  13. NMR determination of pKa values in α-synuclein.
    Protein Sci. 2011 Feb;20(2):256-69 PMID: 21280118
  14. Gaucher disease: gene frequencies and genotype/phenotype correlations.
    Genet Test. 1997;1(1):5-12 PMID: 10464619
  15. Structure of acid beta-glucosidase with pharmacological chaperone provides insight into Gaucher disease.
    Nat Chem Biol. 2007 Feb;3(2):101-7 PMID: 17187079
  16. Biophysics of Parkinson's disease: structure and aggregation of alpha-synuclein.
    Curr Protein Pept Sci. 2009 Oct;10(5):483-99 PMID: 19538146
  17. Interaction of saposins, acidic lipids, and glucosylceramidase.
    J Biol Chem. 1990 Feb 5;265(4):1933-7 PMID: 2298731
  18. X-ray structure of human acid-beta-glucosidase covalently bound to conduritol-B-epoxide. Implications for Gaucher disease.
    J Biol Chem. 2005 Jun 24;280(25):23815-9 PMID: 15817452
  19. Part II: alpha-synuclein and its molecular pathophysiological role in neurodegenerative disease.
    Neuropharmacology. 2003 Jul;45(1):14-44 PMID: 12814657
  20. Multicenter analysis of glucocerebrosidase mutations in Parkinson's disease.
    N Engl J Med. 2009 Oct 22;361(17):1651-61 PMID: 19846850
  21. Glucocerebrosidase mutations in subjects with parkinsonism.
    Mol Genet Metab. 2004 Jan;81(1):70-3 PMID: 14728994
  22. Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson's disease.
    Nat Genet. 1998 Feb;18(2):106-8 PMID: 9462735
  23. Alpha-synuclein locus duplication as a cause of familial Parkinson's disease.
    Lancet. 2004 Sep 25-Oct 1;364(9440):1167-9 PMID: 15451224
  24. Gaucher disease with parkinsonian manifestations: does glucocerebrosidase deficiency contribute to a vulnerability to parkinsonism?
    Mol Genet Metab. 2003 Jun;79(2):104-9 PMID: 12809640
  25. Expanding insights of mitochondrial dysfunction in Parkinson's disease.
    Nat Rev Neurosci. 2006 Mar;7(3):207-19 PMID: 16495942
  26. Analysis of glucocerebrosidase activity using N-(1-[14C]hexanoyl)-D-erythroglucosylsphingosine demonstrates a correlation between levels of residual enzyme activity and the type of Gaucher disease.
    Biochem J. 1994 Oct 15;303 ( Pt 2):377-82 PMID: 7980395
  27. Molecular crowding accelerates fibrillization of alpha-synuclein: could an increase in the cytoplasmic protein concentration induce Parkinson's disease?
    Biochemistry. 2002 Mar 26;41(12):3855-60 PMID: 11900526
  28. Metal-triggered structural transformations, aggregation, and fibrillation of human alpha-synuclein. A possible molecular NK between Parkinson's disease and heavy metal exposure.
    J Biol Chem. 2001 Nov 23;276(47):44284-96 PMID: 11553618
  29. X-ray structure of human acid-beta-glucosidase, the defective enzyme in Gaucher disease.
    EMBO Rep. 2003 Jul;4(7):704-9 PMID: 12792654
  30. Residue-specific fluorescent probes of α-synuclein: detection of early events at the N- and C-termini during fibril assembly.
    Biochemistry. 2011 Mar 29;50(12):1963-5 PMID: 21338068
  31. Parkinsonism among Gaucher disease carriers.
    J Med Genet. 2004 Dec;41(12):937-40 PMID: 15591280
  32. Analysis of human acid beta-glucosidase by site-directed mutagenesis and heterologous expression.
    J Biol Chem. 1994 Jan 21;269(3):2283-91 PMID: 8294487
  33. Mutations in LRRK2 cause autosomal-dominant parkinsonism with pleomorphic pathology.
    Neuron. 2004 Nov 18;44(4):601-7 PMID: 15541309
  34. NMRPipe: a multidimensional spectral processing system based on UNIX pipes.
    J Biomol NMR. 1995 Nov;6(3):277-93 PMID: 8520220
  35. Alpha-synuclein structures from fluorescence energy-transfer kinetics: implications for the role of the protein in Parkinson's disease.
    Proc Natl Acad Sci U S A. 2004 Nov 23;101(47):16466-71 PMID: 15536128
  36. The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia.
    Ann Neurol. 2004 Feb;55(2):164-73 PMID: 14755719
  37. Mutations in the glucocerebrosidase gene and Parkinson's disease in Ashkenazi Jews.
    N Engl J Med. 2004 Nov 4;351(19):1972-7 PMID: 15525722
  38. Early aggregation steps in alpha-synuclein as measured by FCS and FRET: evidence for a contagious conformational change.
    Biophys J. 2010 Apr 7;98(7):1302-11 PMID: 20371330
  39. Multiparametric fluorescence detection of early stages in the amyloid protein aggregation of pyrene-labeled alpha-synuclein.
    J Mol Biol. 2008 May 16;378(5):1064-73 PMID: 18433772
  40. Alpha-synuclein-glucocerebrosidase interactions in pharmacological Gaucher models: a biological link between Gaucher disease and parkinsonism.
    Neurotoxicology. 2009 Nov;30(6):1127-32 PMID: 19576930
  41. Concentration dependence of alpha-synuclein fibril length assessed by quantitative atomic force microscopy and statistical-mechanical theory.
    Biophys J. 2008 Nov 15;95(10):4871-8 PMID: 18676659
  42. Gaucher disease.
    Curr Opin Chem Biol. 2007 Aug;11(4):412-8 PMID: 17644022
  43. Very fast prediction and rationalization of pKa values for protein-ligand complexes.
    Proteins. 2008 Nov 15;73(3):765-83 PMID: 18498103
  44. Alpha-synuclein promotes SNARE-complex assembly in vivo and in vitro.
    Science. 2010 Sep 24;329(5999):1663-7 PMID: 20798282
  45. Acid beta-glucosidase: insights from structural analysis and relevance to Gaucher disease therapy.
    Biol Chem. 2008 Nov;389(11):1361-9 PMID: 18783340
  46. Copper(II) binding to alpha-synuclein, the Parkinson's protein.
    J Am Chem Soc. 2008 Jun 4;130(22):6898-9 PMID: 18465859
  47. alpha-Synuclein membrane interactions and lipid specificity.
    J Biol Chem. 2000 Nov 3;275(44):34328-34 PMID: 10915790
  48. Conformational properties of alpha-synuclein in its free and lipid-associated states.
    J Mol Biol. 2001 Apr 6;307(4):1061-73 PMID: 11286556
  49. Occurrence of Parkinson's syndrome in type I Gaucher disease.
    QJM. 1996 Sep;89(9):691-4 PMID: 8917744
  50. The aggregation and fibrillation of alpha-synuclein.
    Acc Chem Res. 2006 Sep;39(9):628-34 PMID: 16981679
  51. Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism.
    Nature. 1998 Apr 9;392(6676):605-8 PMID: 9560156
  52. Parkin protects against the toxicity associated with mutant alpha-synuclein: proteasome dysfunction selectively affects catecholaminergic neurons.
    Neuron. 2002 Dec 19;36(6):1007-19 PMID: 12495618
  53. Solution structure of human saposin C: pH-dependent interaction with phospholipid vesicles.
    Biochemistry. 2003 Dec 23;42(50):14729-40 PMID: 14674747
  54. Hereditary early-onset Parkinson's disease caused by mutations in PINK1.
    Science. 2004 May 21;304(5674):1158-60 PMID: 15087508
  55. Emerging pathways in genetic Parkinson's disease: Potential role of ceramide metabolism in Lewy body disease.
    FEBS J. 2008 Dec;275(23):5767-73 PMID: 19021754
  56. Mechanisms of Parkinson's disease linked to pathological alpha-synuclein: new targets for drug discovery.
    Neuron. 2006 Oct 5;52(1):33-8 PMID: 17015225
  57. Protein misfolding, functional amyloid, and human disease.
    Annu Rev Biochem. 2006;75:333-66 PMID: 16756495
  58. Accelerated in vitro fibril formation by a mutant alpha-synuclein linked to early-onset Parkinson disease.
    Nat Med. 1998 Nov;4(11):1318-20 PMID: 9809558
  59. Synuclein: a neuron-specific protein localized to the nucleus and presynaptic nerve terminal.
    J Neurosci. 1988 Aug;8(8):2804-15 PMID: 3411354
  60. Cathepsin D: a cellular roadmap.
    Biochem Biophys Res Commun. 2008 Nov 7;376(1):5-9 PMID: 18762174
  61. Identification of two distinct synucleins from human brain.
    FEBS Lett. 1994 May 23;345(1):27-32 PMID: 8194594
  62. Do amyloid oligomers act as traps for misfolded proteins? A hypothesis.
    Amyloid. 2008 Sep;15(3):160-5 PMID: 18925454
  63. The synucleins: a family of proteins involved in synaptic function, plasticity, neurodegeneration and disease.
    Trends Neurosci. 1998 Jun;21(6):249-54 PMID: 9641537
  64. Glucocerebrosidase is present in α-synuclein inclusions in Lewy body disorders.
    Acta Neuropathol. 2010 Nov;120(5):641-9 PMID: 20838799
  65. Lysosomal degradation of alpha-synuclein in vivo.
    J Biol Chem. 2010 Apr 30;285(18):13621-9 PMID: 20200163
  66. The role of glucocerebrosidase mutations in Parkinson disease and Lewy body disorders.
    Curr Neurol Neurosci Rep. 2010 May;10(3):190-8 PMID: 20425034
  67. Gaucher disease and parkinsonism, a molecular link theory.
    Mol Genet Metab. 2010 Dec;101(4):307-10 PMID: 20801700
  68. Solution structure of human saposin C in a detergent environment.
    J Mol Biol. 2005 Mar 11;346(5):1381-92 PMID: 15713488
  69. Gaucher disease: mutation and polymorphism spectrum in the glucocerebrosidase gene (GBA).
    Hum Mutat. 2008 May;29(5):567-83 PMID: 18338393
  70. Multiple tight phospholipid-binding modes of alpha-synuclein revealed by solution NMR spectroscopy.
    J Mol Biol. 2009 Jul 24;390(4):775-90 PMID: 19481095
  71. The lipid-binding domain of wild type and mutant alpha-synuclein: compactness and interconversion between the broken and extended helix forms.
    J Biol Chem. 2010 Sep 3;285(36):28261-74 PMID: 20592036
  72. Tryptophan probes at the alpha-synuclein and membrane interface.
    J Phys Chem B. 2010 Apr 8;114(13):4615-22 PMID: 20229987
  73. Small molecule inhibitors of alpha-synuclein filament assembly.
    Biochemistry. 2006 May 16;45(19):6085-94 PMID: 16681381
  74. A block of autophagy in lysosomal storage disorders.
    Hum Mol Genet. 2008 Jan 1;17(1):119-29 PMID: 17913701
  75. X-ray and biochemical analysis of N370S mutant human acid β-glucosidase.
    J Biol Chem. 2011 Jan 7;286(1):299-308 PMID: 20980263
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
Published
2011-08-12
Epub
2011-00-08
Pages
28080-8
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3151053
Subset
IM
Grants
Intramural NIH HHS · ZIA HG200336-06 · United States
Intramural NIH HHS · ZIA HL001055-04 · United States
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