[en] Alpha-synuclein is a 140 residue protein associated with Parkinson's disease. Intraneural inclusions called Lewy bodies and Lewy neurites are mainly composed of alpha-synuclein aggregated into amyloid fibrils. Other amyloidogenic proteins, such as the beta amyloid peptide involved in Alzheimer's disease and the prion protein (PrP) associated with Creuztfeldt-Jakob's disease, are known to possess "tilted peptides". These peptides are short protein fragments that adopt an oblique orientation at a hydrophobic/hydrophilic interface, which enables destabilization of the membranes. In this paper, sequence analysis and molecular modelling predict that the 67-78 fragment of alpha-synuclein is a tilted peptide. Its destabilizing properties were tested experimentally. The alpha-synuclein 67-78 peptide is able to induce lipid mixing and leakage of unilamellar liposomes. The neuronal toxicity, studied using human neuroblastoma cells, demonstrated that the alpha-synuclein 67-78 peptide induces neurotoxicity. A mutant designed by molecular modelling to be amphipathic was shown to be significantly less fusogenic and toxic than the wild type. In conclusion, we have identified a tilted peptide in alpha-synuclein, which could be involved in the toxicity induced during amyloidogenesis of alpha-synuclein.
Maroteaux L, Campanelli JT, Scheller RH. Synuclein: a neuron-specific protein localized to the nucleus and presynaptic nerve terminal. J Neurosci 1988;8:2804-2815.
Jakes R, Spillantini MG, Goedert M. Identification of two distinct synucleins from human brain. FEBS Lett 1994;345:27-32.
Spillantini MG, Crowther RA, Jakes R, Hasegawa M, Goedert M. α-synuclein in filamentous inclusions of Lewy bodies from Parkinson's disease and dementia with lewy bodies. Proc Natl Acad Sci USA 1998;95:6469-6473.
Spillantini MG, Crowther RA, Jakes R, Cairns NJ, Lantos PL, Goedert M. Filamentous α-synuclein inclusions link multiple system atrophy with Parkinson's disease and dementia with Lewy bodies. Neurosci Lett 1998;251:205-208.
Weinreb PH, Zhen W, Poon AW, Conway KA, Lansbury PT, Jr. NACP, a protein implicated in Alzheimer's disease and learning, is natively unfolded. Biochemistry 1996;35:13709-13715.
Davidson WS, Jonas A, Clayton DF, George JM. Stabilization of α-synuclein secondary structure upon binding to synthetic membranes. J Biol Chem 1998;273:9443-9449.
Ulmer TS, Bax A, Cole NB, Nussbaum RL. Structure and dynamics of micelle-bound human α-synuclein. J Biol Chem 2005;280:9595-9603.
Bussell R, Eliezer D. A structural and functional role for 11-mer repeats in α-synuclein and other exchangeable lipid binding proteins. J Mol Biol 2003;329:763-778.
Zhu M, Li J, Fink AL. The association of α-synuclein with membranes affects bilayer structure, stability, and fibril formation. J Biol Chem 2003;278:40186-40197.
Miake H, Mizusawa H, Iwatsubo T, Hasewaga M. Biochemical characterization of the core structure of α-synuclein filaments. J Biol Chem 2002;277:19213-19219.
Del Mar C, Geenbaum EA, Mayne L, Englander SW, Woods V. Structure and properties of α-synuclein and other amyloids determined at the amino acid level. Proc Natl Acad Sci USA 2005;102: 15477-15482.
Ueda K, Fukushima H, Masliah E, Xia Y, Iwai A, Yoshimoto M, Otero DA, Kondo J, Ihara Y, Saitoh T. Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease. Proc Natl Acad Sci USA 1993;90:11282-11286.
Iwai A. Properties of NACP/α-synuclein and its role in Alzheimer's disease. Biochim Biophys Acta 2000;1502:95-109.
Giasson BI, Murray IV, Trojanowski JQ, Lee VM. A hydrophobic stretch of 12 amino acid residues in the middle of α-synuclein is essential for filament assembly. J Biol Chem 2001;276:2380-2386.
El-Agnaf OMA, Bodles A, Guthrie DJS, Harriott P, Irvine GB. The N-terminal region of non-Aβ component of Alzheimer's Disease amyloid is responsible for its tendency to assume β-sheet and aggregate to form fibrils. Eur J Biochem 1998;258:157-163.
El-Agnaf OMA, Irvine GB. Aggregation and neurotoxicity of α-synuclein and related peptides. Biochem Soc Trans 2002;30:559-565.
Pillot T, Goethals M, Vanloo B, Talussot C, Brasseur R, Vandekerckhove J, Rosseneu M, Lins L. Fusogenic properties of the C-terminal domain of the Alzheimer β-amyloid peptide. J Biol Chem 1996;271:28757-28765.
Pillot T, Lins L, Goethals M, Vanloo B, Baert J, Vandekerckhove J, Rosseneu M, Brasseur R. The 118-135 peptide of the human prion protein forms amyloid fibrils and induces liposome fusion. J Mol Biol 1997;274:381-393.
Lins L, Charloteaux B, Thomas A, Brasseur R. Computational study of lipid-destabilizing protein fragments: towards a comprehensive view of tilted peptides. Proteins 2001;44:435-447.
Brasseur R. Tilted peptides: a motif for membrane destabilization (Hypothesis). Mol Membr Biol 2000;17:31-40.
Colotto A, Martin I, Ruysschaert JM, Sen A, Flui SW, Epand RM. Structural study of the interaction between the SIV fusion peptide and model membranes. Biochemistry 1996;35:980-989.
Pérez-Méndez O, Vanloo B, Decout A, Goethals M, Peelman F, Vandekerckhove J, Brasseur R, Rosseneu M. Contribution of the hydrophobicity gradient of an amphipathic peptide to its mode of association with lipids. Eur J Biochem 1998;256:570-579.
Horth M, Lambrecht B, Khim MCL, Bex F, Thiriart C, Ruysschaert JM, Burny A, Brasseur R. Theorical and functional analysis of the SIV fusion peptide. EMBO J 1991;10:2747-2755.
Martin I, Dubois MC, Defrise-Quertain F, Saermark T, Burny A, Brasseur R, Ruysschaert JM. Correlation between fusogenicity of synthetic modified peptides corresponding to the NH2-terminal extremity of simian immunodeficiency virus gp32 and their mode of insertion into the lipid bilayer: an infrared spectroscopy study. J Virol 1994;68:1139-1148.
Charloteaux B, Lorin A, Crowet JM, Stroobant V, Lins L, Thomas A, Brasseur R. The N-terminal 12 residue long peptide of HIV gp41 is the minimal peptide sufficient to induce significant T-cell-like membrane destabilization in vitro. J Mol Biol 2006;359:597-609.
Lorin A, Thomas A, Stroobant V, Brasseur R, Lins L. Lipid-destabilising properties of a peptide with structural plasticity. Chem Phys Lipids 2006;141:185-196.
Martin I, Schaal H, Scheid A, Ruysschaert JM. Lipid membrane fusion induced by the human immunodeficiency virus type I gp41 N-terminal extremity is determined by its orientation in the lipid bilayer. J Virol 1996;70:298-304.
Castano S, Desbat B. Structure and orientation study of fusion peptide FP23 of gp41 from HIV-1 alone or inserted into various lipid membrane models (mono-, bi- and multibi-layers) by FT-IR spectroscopies and Brewster angle microscopy. Biochim Biophys Acta 2005;1715:81-95.
Bradshaw JP, Darkes MJ, Harroun TA, Katsaras J, Epand RM. Oblique membrane insertion of viral fusion peptide probed by neutron diffraction. Biochemistry 2000;39:6581-6585.
Han X, Bushweller JH, Cafiso DS, Tamm LK. Membrane structure and fusion-triggering conformational change of the fusion domain from influenza hemagglutinin. Nat Struct Biol 2001;8:715-720.
Brasseur R, Pillot T, Lins L, Vandekerckhove I, Rosseneu M. Peptides in membranes: tipping the balance of membrane stability. Trends Biochem Sci 1997;22:167-171.
Lansbury PTJ, Costa JM, Griffiths EJ, Simon EJ, Auger M, Halverson KJ, Kocisko DA, Hendsch ZS, Ashburn TT, Spencer RG. Structural model for the β-amyloid fibril based on interstrand alignment of an antiparallel-sheet comprising a C-terminal peptide. Nat Struct Biol 1995;2:990-998.
Nguyen J, Baldwin MA, Cohen FE, Prusiner SB. Prion protein peptides induce α-helix to β-sheet conformational transitions. Biochemistry 1995;34:4186-4192.
Gaboriaud C, Bissery V, Benchetrit T, Mornon JP. Hydrophobic cluster analysis an efficient new way to compare and analyse amino acid sequence. FEBS Lett 1987;224:149-155.
Jahnig F. Structure predictions of membrane proteins are not that bad. Trends Biochem Sci 1990;15:93-95.
Brasseur R, Lins L, Vanloo B, Ruysschaert JM, Rosseneu M. Molecular modelling of the amphipathic helices of the plasma apolipoproteins. Proteins 1992;13:246-257.
Ducarme P, Rahman M, Brasseur R. Impala: a simple restrain: field to simulate the biological membrane in molecular structure studies. Proteins 1998;30:357-371.
Lins L, Charloteaux B, Heinen C, Thomas A, Brasseur R. "De novo" design of peptides with specific lipid-binding properties. Biophys J 2006;90:470-479.
Brasseur R. Simulating the folding of small proteins by use of the local minimum energy and the free solvation energy yields native-like structures. J Mol Graph 1995;13:312-322.
Thomas A, Desbayes S, Deccaffmeyer M, Van Eyck MH, Charloteaux B, Brasseur R. Prediction of peptide structure: How far are we? Proteins 2006;65:889-897.
Etchebest C, Benros C, Hazout S, de Brevern AG. A structural alphabet for local protein structure: improved prediction methods. Proteins 2005;59:810-827.
Nelder JA, Mead RA. A simplex method for function minimization. Comput J 1997;7:303-313.
Mayer LD, Hope MJ, Cullis PR. Vesicles of variable sizes produced by a rapid extrusion procedure. Biochim Biophys Acta 1986;858: 161-168.
Mrsny RJ, Volwerk JJ, Griffith OH. A simplified procedure for lipid phosphorus analysis shows that digestion rates vary with phospholipid structure. Chem Phys Lipids 1986;39:185-191.
Hoekstra D, De Boer T, Klappe K, Wilschut J. Fluorescence method for measuring the kinetics of fusion between biological membranes. Biochemistry 1984;23:5675-5681.
Ellens H, Bentz J, Szoka FC. H+- and Ca2+-induced fusion and destabilization of liposomes. Biochemistry 1985;24:3099-3106.
Sreerama N, Woody RW. Estimation of protein secondary structure from CD spectra: comparison of CONTIN, SELCON and CDSSTR methods with an expanded reference set. Anal Biochem 2000;287: 252-260.
Eisenberg D, Weiss R, Terwilliger T. The helical hydrophobic moment: a measure of the amphiphilicity of the α-helix. Nature 1982;299:371-374.
Jonson PH, Petersen SB. A critical view on conservative mutations. Protein Eng 2001;14:397-402.
Hofmann MW, Weise K, Ollesch J, Agrawal P, Stalz H, Stelzer W, Hulsbergen F, de Groot H, Gerwert K, Reed J, Langosch D. De novo design of conformationally flexible transmembrane peptides driving membrane fusion. Proc Natl Acad Sci USA 2004;101:14776-14781.
Brasseur R, Lorge P, Espion D, Goormaghtigh E, Burny A, Ruysschaert JM. The Mode of insertion of the paramyxovirus F1 N-terminus into lipid matrix, an initial step in host cell-virus fusion. Virus Genes 1988;1:325-332.
Vonèche V, Portetelle D, Kettman R, Willems L, Limbach K, Paoletti E, Ruysschaert JM, Burny A, Brasseur R. Fusogenic segment of bovine leucemia virus and simian immunodeficiency virus are interchangeable and mediate fusion by means of oblique insertion in the lipid bilayer of their target cells. Proc Natl Acad Sci USA 1992;89:3810-3814.
Du HN, Tang L, Luo XY, Li HT, Hu J, Zhou JW, Hu HY. A peptide motif consisting of glycine, alanine, and valine is required for the fibrillization and cytotoxicity of human α-synuclein. Biochemistry 2003;42:8870-8878.
Brasseur R, Ruysschaert JM. Conformation and mode of organiza-tion of amphiphilic membrane components: a conformational analysis. Biochem J 1986;238:1-11.
Litis L, El Kirat K, Flore C, Stroobant V, Thomas A, Brasseur R. Lipid-destabilizing properties of the hydrophobic helices F18 and F19 from colicin E1. Mol Membr Biol, in press.
Conway KA, Lee SJ, Rochet JC, Ding TT, Williamson RE, Lansbury PT. Acceleration of oligomerization, not fibrillization, is a shared property of both α-synuclein mutations linked to early-onset Parkinson's disease: implications for pathogenesis and therapy. Proc Natl Acad Sci USA 2000;97:571-576.
Goldberg MS, Lansbury PT, Jr. Is there a cause-and-effect relationship lietween α-synuclein fibrillization and Parkinson's disease? Nat Cell Biol 2000;2:115-119.
Lashuel HA, Hartley D, Petre BM, Walz T, Lansbury PT. Neurodegenerative disease: amyloid pores from pathogenic mutations. Nature 2002;418:291-291.
Volles MJ, Lansbury PT. Vesicle permeabilization by protofibrillar α-synuclein is sensitive to Parkinson's disease-linked mutations and occurs by a pore-like mechanism. Biochemistry 2002;41:4595-4602.
Pountney DL, Lowe R, Quilty M, Vickers JC, Voelcker NH, Gai WP. Annular α-synuclein species from purified multiple system atrophy inclusions. J Neurochem 2004;90:502-512.
Lins L, Charloteaux B, Thomas A, Brasseur R. Implication of a structural motif in the instability of a toxic protein: the Prion. In: Renaville R, Burny A, editors. Biotechnology in animal husbandry. Boston, MA: Kluwer; 2001. pp 15-32.
Quist A, Doudevski I, Lin H, Azimova R, Ng D, Frangione B, Kagan B, Ghiso J, Lai R. Amyloid ion channels: a common structural link for protein-misfolding disease. Proc Natl Acad Sci USA 2005;102:10427-10432.
Lin H, Bhatia R, Lal R. Amyloid B protein forms ion channels: implications for Alzheimer's disease pathophysiology. FASEB J 2001;15:2433-2444.
Apetri MM, Maiti NC, Zagorski MG, Carey PR, Anderson VE. Secondary structure of α-synuclein oligomers: characterization by raman and atomic force microscopy. J Mol Biol 2006;355:63-71.
Kirkitadze MD, Condron MM, Teplow DB. Identification and characterization of key kinetic intermediates in amyloid β-protein fibrillogenesis. J Mol Biol 2001;312:1103-1119.
Crowther RA, Jakes R, Spillantini MG, Goedert M. Synthetic filaments assembled from C-terminally truncated α-synuclein. FEBS Lett 1998;436:309-312.
Lins L, Thomas-Soumarmon A, Pillot T, Vandekerckhove J, Rosseneu M, Brasseur R. Molecular determinants of the interaction between the C-terminal domain of Alzheimer's β-amyloid peptide and apolipoprotein E α-helices. J Neurochem 1999;73:758-769.
Pillot T, Goethals M, Najib J, Labeur C, Lins L, Chambaz J, Brasseur R, Vandekerckhove J, Rosseneu M. β-amyloid peptide interacts specifically with the carboxy-terminal domain of human apolipoprotein E: relevance to Alzheimer's disease. J Neurochem 1999; 72:230-237.
Decaffmeyer M, Lins L, Charloteaux B, Van Eyck MH, Thomas A, Brasseur R. Rational design of complementary peptides to the βAmyloid 29-42 fusion peptide: an application of PepDesign. Biochim Biophys Acta 2006;1758:320-327.
