Rapid Anionic Micelle-mediated α-Synuclein Fibrillization in Vitro*

Parkinson's disease is characterized by the aggregation of α-synuclein into filamentous forms within affected neurons of the basal ganglia. Fibrillization of purified recombinant α-synuclein is inefficient in vitro but can be enhanced by the addition of various agents including glycosaminoglycans and polycations. Here we report that fatty acids and structurally related anionic detergents greatly accelerate fibrillization of recombinant α-synuclein at low micromolar concentrations with lag times as short as 11 min and apparent first order growth rate constants as fast as 10.4 h–1. All detergents and fatty acids were micellar at active concentrations because of an α-synuclein-dependent depression of their critical micelle concentrations. Other anionic surfaces, such as those supplied by anionic phospholipid vesicles, also induced α-synuclein fibrillization, with resultant filaments originating from their surface. These data suggest that anionic surfaces presented as micelles or vesicles can serve to nucleate α-synuclein fibrillization, that this mechanism underlies the inducer activity of anionic surfactants, and that anionic membranes may serve this function in vivo.

[1]  Akihiko Iwai,et al.  The precursor protein of non-Aβ component of Alzheimer's disease amyloid is a presynaptic protein of the central nervous system , 1995, Neuron.

[2]  M. Hosokawa,et al.  Fluorometric determination of amyloid fibrils in vitro using the fluorescent dye, thioflavin T1. , 1989, Analytical biochemistry.

[3]  Olaf Riess,et al.  AlaSOPro mutation in the gene encoding α-synuclein in Parkinson's disease , 1998, Nature Genetics.

[4]  Yamabe,et al.  Micelle Formation of Anionic Surfactant with Divalent Counterion of Separate Electric Charge. , 1999, Journal of colloid and interface science.

[5]  P. Lansbury,et al.  Fibrils formed in vitro from alpha-synuclein and two mutant forms linked to Parkinson's disease are typical amyloid. , 2000, Biochemistry.

[6]  C. Bergeron,et al.  Lewy Bodies in Alzheimer Disease‐One or Two Diseases? , 1989, Alzheimer disease and associated disorders.

[7]  Peter T. Lansbury,et al.  Accelerated in vitro fibril formation by a mutant α-synuclein linked to early-onset Parkinson disease , 1998, Nature Medicine.

[8]  R. Crowther,et al.  α-Synuclein in filamentous inclusions of Lewy bodies from Parkinson’s disease and dementia with Lewy bodies , 1998 .

[9]  G. Withers,et al.  Delayed localization of synelfin (synuclein, NACP) to presynaptic terminals in cultured rat hippocampal neurons. , 1997, Brain research. Developmental brain research.

[10]  Shah,et al.  Prediction of Critical Micelle Concentration Using a Quantitative Structure-Property Relationship Approach , 1997, Journal of colloid and interface science.

[11]  P. Blumbergs,et al.  In Situ and in Vitro Study of Colocalization and Segregation of α-Synuclein, Ubiquitin, and Lipids in Lewy Bodies , 2000, Experimental Neurology.

[12]  M. L. Schmidt,et al.  α-Synuclein in Lewy bodies , 1997, Nature.

[13]  D. Eliezer,et al.  Conformational properties of alpha-synuclein in its free and lipid-associated states. , 2001, Journal of molecular biology.

[14]  M. Goedert,et al.  Binding of α-Synuclein to Brain Vesicles Is Abolished by Familial Parkinson’s Disease Mutation* , 1998, The Journal of Biological Chemistry.

[15]  B. Bales.  A Definition of the Degree of Ionization of a Micelle Based on Its Aggregation Number , 2001 .

[16]  C. Chirita,et al.  Anionic Micelles and Vesicles Induce Tau Fibrillization in Vitro* , 2003, Journal of Biological Chemistry.

[17]  G. Asano,et al.  α-Synuclein-positive structures in cases with sporadic Alzheimer’s disease: morphology and its relationship to tau aggregation , 2001, Brain Research.

[18]  Robert L. Nussbaum,et al.  Mutation in the α-Synuclein Gene Identified in Families with Parkinson's Disease , 1997 .

[19]  C. Dobson,et al.  Stimulation and inhibition of fibril formation by a peptide in the presence of different concentrations of SDS , 2002, FEBS letters.

[20]  B. Bales,et al.  Surfactant- and Salt-Induced Growth of Normal Sodium Alkyl Sulfate Micelles Well above Their Critical Micelle Concentrations , 2000 .

[21]  P. Lansbury,et al.  Seeding “one-dimensional crystallization” of amyloid: A pathogenic mechanism in Alzheimer's disease and scrapie? , 1993, Cell.

[22]  R. Anthony Crowther,et al.  Synthetic filaments assembled from C‐terminally truncated α‐synuclein , 1998 .

[23]  L. Mucke,et al.  Dopaminergic loss and inclusion body formation in alpha-synuclein mice: implications for neurodegenerative disorders. , 2000, Science.

[24]  P. Lansbury,et al.  Acceleration of oligomerization, not fibrillization, is a shared property of both alpha-synuclein mutations linked to early-onset Parkinson's disease: implications for pathogenesis and therapy. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[25]  V. Uversky,et al.  Evidence for a Partially Folded Intermediate in α-Synuclein Fibril Formation* , 2001, The Journal of Biological Chemistry.

[26]  Min Zhu,et al.  Lipid Binding Inhibits α-Synuclein Fibril Formation* , 2003, The Journal of Biological Chemistry.

[27]  J. Kuret,et al.  Ligand-dependent tau filament formation: implications for Alzheimer's disease progression. , 1999, Biochemistry.

[28]  D. Dickson,et al.  Pathology and Biology of the Lewy Body , 1993, Journal of neuropathology and experimental neurology.

[29]  V. Uversky,et al.  Heparin and other glycosaminoglycans stimulate the formation of amyloid fibrils from alpha-synuclein in vitro. , 2002, Biochemistry.

[30]  M. Kirkitadze,et al.  Identification and characterization of key kinetic intermediates in amyloid beta-protein fibrillogenesis. , 2001, Journal of molecular biology.

[31]  K. Arima,et al.  Tubulin Seeds α-Synuclein Fibril Formation* , 2002, The Journal of Biological Chemistry.

[32]  M. Hoshino,et al.  Aggregation of beta(2)-glycoprotein I induced by sodium lauryl sulfate and lysophospholipids. , 2002, Biochemistry.

[33]  W. Vaz,et al.  Determination of the critical micelle concentration of surfactants using the fluorescent probe N-phenyl-1-naphthylamine. , 1986, Analytical biochemistry.

[34]  J Q Trojanowski,et al.  Lewy bodies contain altered alpha-synuclein in brains of many familial Alzheimer's disease patients with mutations in presenilin and amyloid precursor protein genes. , 1998, The American journal of pathology.

[35]  P. S. St George-Hyslop,et al.  α-Synuclein Membrane Interactions and Lipid Specificity* , 2000, The Journal of Biological Chemistry.

[36]  W. Honer,et al.  α‐Synuclein Inclusions in Alzheimer and Lewy Body Diseases , 2000, Journal of neuropathology and experimental neurology.

[37]  W. Honer,et al.  Abnormal accumulation of NACP/alpha-synuclein in neurodegenerative disorders. , 1998, The American journal of pathology.

[38]  V. Uversky,et al.  Metal-triggered structural transformations, aggregation, and fibrillation of human alpha-synuclein. A possible molecular NK between Parkinson's disease and heavy metal exposure. , 2001, The Journal of biological chemistry.

[39]  P. Lansbury,et al.  Vesicle permeabilization by protofibrillar alpha-synuclein: implications for the pathogenesis and treatment of Parkinson's disease. , 2001, Biochemistry.

[40]  S. Love,et al.  Microtubule-associated protein tau, heparan sulphate and α-synuclein in several neurodegenerative diseases with dementia , 1999, Acta Neuropathologica.

[41]  R. Scheller,et al.  Synuclein: a neuron-specific protein localized to the nucleus and presynaptic nerve terminal , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[42]  Seung-Jae Lee,et al.  Membrane-bound α-Synuclein Has a High Aggregation Propensity and the Ability to Seed the Aggregation of the Cytosolic Form* , 2002, The Journal of Biological Chemistry.

[43]  R. Jakes,et al.  Effects of the mutations Ala30 to Pro and Ala53 to Thr on the physical and morphological properties of α‐synuclein protein implicated in Parkinson's disease , 1998, FEBS letters.

[44]  S. W. Davies,et al.  Filamentous nerve cell inclusions in neurodegenerative diseases , 1998, Current Opinion in Neurobiology.

[45]  Rebecca A. Betensky,et al.  α-Synuclein occurs in lipid-rich high molecular weight complexes, binds fatty acids, and shows homology to the fatty acid-binding proteins , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[46]  D. Wilson,et al.  Free fatty acids stimulate the polymerization of tau and amyloid beta peptides. In vitro evidence for a common effector of pathogenesis in Alzheimer's disease. , 1997, The American journal of pathology.

[47]  G. J. Raymond,et al.  Conformational change, aggregation and fibril formation induced by detergent treatments of cellular prion protein , 2001, Journal of neurochemistry.

[48]  R. Perrin,et al.  Exposure to Long Chain Polyunsaturated Fatty Acids Triggers Rapid Multimerization of Synucleins* , 2001, The Journal of Biological Chemistry.

[49]  J. Lahti,et al.  Use of gene knockouts in cultured cells to study apoptosis. , 1999, Methods.

[50]  V. Subramaniam,et al.  Cellular Polyamines Promote the Aggregation of α-Synuclein* , 2003, The Journal of Biological Chemistry.

[51]  John Q. Trojanowski,et al.  Neurodegenerative dementias : clinical features and pathological mechanisms , 2000 .

[52]  G. Gunnarsson,et al.  Surfactant association into micelles: an electrostatic approach , 1980 .

[53]  S. Mammi,et al.  Conformation and interactions of uteroglobin fragments 4–14 and 49–65 in aqueous solution containing surfactant micelles , 1993, Biopolymers.

[54]  L. Serpell,et al.  Fiber diffraction of synthetic alpha-synuclein filaments shows amyloid-like cross-beta conformation. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[55]  D. Marsh,et al.  Head group and chain length dependence of phospholipid self-assembly studied by spin-label electron spin resonance. , 1987, Biochemistry.

[56]  C M Dobson,et al.  Formation of insulin amyloid fibrils followed by FTIR simultaneously with CD and electron microscopy , 2000, Protein science : a publication of the Protein Society.

[57]  J. Trojanowski,et al.  Antibodies to α‐synuclein detect Lewy bodies in many Down's syndrome brains with Alzheimer's disease , 1999, Annals of neurology.

[58]  S. Nakajo,et al.  Cell and tissue distribution and developmental change of neuron specific 14 kDa protein (phosphoneuroprotein 14) , 1993, Brain Research.

[59]  A. Jonas,et al.  Stabilization of α-Synuclein Secondary Structure upon Binding to Synthetic Membranes* , 1998, The Journal of Biological Chemistry.

[60]  M. Citron,et al.  alpha-synuclein fibrillogenesis is nucleation-dependent. Implications for the pathogenesis of Parkinson's disease. , 1999, The Journal of biological chemistry.

[61]  Nigel J. Cairns,et al.  Filamentous α-synuclein inclusions link multiple system atrophy with Parkinson's disease and dementia with Lewy bodies , 1998, Neuroscience Letters.

[62]  B. Hyman,et al.  Characterization of the Precursor Protein of the Non-Aβ Component of Senile Plaques (NACP) in the Human Central Nervous System , 1996, Journal of neuropathology and experimental neurology.

[63]  J. Trojanowski,et al.  Mutant and Wild Type Human α-Synucleins Assemble into Elongated Filaments with Distinct Morphologies in Vitro * , 1999, The Journal of Biological Chemistry.

[64]  P J McLean,et al.  Membrane association and protein conformation of alpha-synuclein in intact neurons. Effect of Parkinson's disease-linked mutations. , 2000, The Journal of biological chemistry.

[65]  J. Trojanowski,et al.  “Fatal Attractions” of Proteins: A Comprehensive Hypothetical Mechanism Underlying Alzheimer's Disease and Other Neurodegenerative Disorders , 2000, Annals of the New York Academy of Sciences.

[66]  V. Tennyson,et al.  Phase and Electron Microscopic Observations of Lewy Bodies and Melanin Granules in the Substantia Nigra and Locus Caeruleus in Parkinson's Disease , 1965 .

[67]  J. Kuret,et al.  The Structural Basis of Monoclonal Antibody Alz50's Selectivity for Alzheimer's Disease Pathology* , 1996, The Journal of Biological Chemistry.

[68]  M. Polymeropoulos,et al.  Mapping of a Gene for Parkinson's Disease to Chromosome 4q21-q23 , 1996, Science.

[69]  W. D. Harkins,et al.  The effect of salts on the critical concentration for the formation of micelles in colloidal electrolytes. , 1947, Journal of the American Chemical Society.

[70]  Christian Haass,et al.  Subcellular Localization of Wild-Type and Parkinson's Disease-Associated Mutant α-Synuclein in Human and Transgenic Mouse Brain , 2000, The Journal of Neuroscience.

[71]  S. Murayama,et al.  Lewy body in neurodegeneration with brain iron accumulation type 1 is immunoreactive for α-synuclein , 1998, Neurology.

[72]  P. S. St George-Hyslop,et al.  Defective membrane interactions of familial Parkinson's disease mutant A30P α-synuclein , 2002 .

[73]  V. Uversky,et al.  Effect of environmental factors on the kinetics of insulin fibril formation: elucidation of the molecular mechanism. , 2001, Biochemistry.

[74]  C. Ionescu-Zanetti,et al.  Surface-catalyzed Amyloid Fibril Formation* , 2002, The Journal of Biological Chemistry.

[75]  M. Citron,et al.  Both Familial Parkinson’s Disease Mutations Accelerate α-Synuclein Aggregation* , 1999, The Journal of Biological Chemistry.

[76]  W. A. Jager,et al.  Sphingomyelin in Lewy inclusion bodies in Parkinson's disease. , 1969 .

[77]  P. Lansbury,et al.  Apolipoprotein E is a kinetic but not a thermodynamic inhibitor of amyloid formation: implications for the pathogenesis and treatment of Alzheimer disease. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[78]  J Q Trojanowski,et al.  Aggregation of alpha-synuclein in Lewy bodies of sporadic Parkinson's disease and dementia with Lewy bodies. , 1998, The American journal of pathology.

[79]  F. Gejyo,et al.  Apolipoprotein E and antioxidants have different mechanisms of inhibiting Alzheimer's beta-amyloid fibril formation in vitro. , 1998, Biochemistry.

[80]  D. Selkoe,et al.  A de novo designed helix-turn-helix peptide forms nontoxic amyloid fibrils , 2000, Nature Structural Biology.