Artículo

La versión final de este artículo es de uso interno de la institución.
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

The aggregation of α-synuclein, a presynaptic protein, has an important role in the etiology of Parkinson's disease. Oligomers or protofibrils adopting the cross-β-sheet structure characteristic of fibrillating amyloid proteins are presumed to be the primary cytotoxic species. Current techniques for monitoring the kinetics of α-synuclein aggregation based on fluorescent dyes such as Thioflavin-T and Congo red detect only the terminal fibrillar species, are discontinuous and notoriously irreproducible. We have devised a new fluorescence aggregation assay that is continuous and provides a large set of fluorescence parameters sensitive to the presence of oligomeric intermediates as well as fibrils. The approach involves tagging functionally neutral Ala-to-Cys variants of α-synuclein with the long-lifetime fluorophore pyrene. Upon induction of aggregation at 37 °C, the entire family of steady-state descriptors of pyrene emission (monomer intensity, solvent polarity ratio (II/IIII), and anisotropy; and excimer intensity) change dramatically, particularly during the early stages in which oligomeric intermediates form and evolve. The pyrene probe senses a progressive decrease in polarity, an increase in molecular mass and close intermolecular association in a manner dependent on position in the sequence and the presence of point mutations. The time-resolved decays (0-160 ns) of intensity and anisotropy exhibited complex, characteristic features. The new assay constitutes a convenient platform for the high-throughput screening of agents useful in the diagnosis and therapy of Parkinson's disease as well as in basic investigations. © 2008 Elsevier Ltd. All rights reserved.

Registro:

Documento: Artículo
Título:Multiparametric Fluorescence Detection of Early Stages in the Amyloid Protein Aggregation of Pyrene-labeled α-Synuclein
Autor:Thirunavukkuarasu, S.; Jares-Erijman, E.A.; Jovin, T.M.
Filiación:Laboratory of Cellular Dynamics, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany
Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, 1428 Buenos Aires, Argentina
Palabras clave:fibrillization; flourescence anisotropy; neurodegenerative; oligomers; Parkinson's disease; alpha synuclein; pyrene; anisotropy; article; fluorescence; human; in vivo study; kinetics; Parkinson disease; point mutation; priority journal; protein aggregation; protein analysis; alpha-Synuclein; Amino Acid Sequence; Amyloid; Anisotropy; Fluorescent Dyes; Humans; Molecular Sequence Data; Mutagenesis, Site-Directed; Parkinson Disease; Protein Folding; Protein Structure, Quaternary; Pyrenes; Spectrometry, Fluorescence
Año:2008
Volumen:378
Número:5
Página de inicio:1064
Página de fin:1073
DOI: http://dx.doi.org/10.1016/j.jmb.2008.03.034
Título revista:Journal of Molecular Biology
Título revista abreviado:J. Mol. Biol.
ISSN:00222836
CODEN:JMOBA
CAS:alpha synuclein, 154040-18-3; pyrene, 129-00-0; alpha-Synuclein; Amyloid; Fluorescent Dyes; pyrene, 129-00-0; Pyrenes
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00222836_v378_n5_p1064_Thirunavukkuarasu

Referencias:

  • Chiti, F., Dobson, C.M., Protein misfolding, functional amyloid, and human disease (2006) Annu. Rev. Biochem., 75, pp. 333-366
  • Hamley, I.W., Peptide fibrillization (2007) Angew Chem. Int. Ed. Engl., 46, pp. 8128-8147
  • Hoyer, W., Cherny, D., Subramaniam, V., Jovin, T.M., Impact of the acidic C-terminal region comprising amino acids 109-140 on α-synuclein aggregation in vitro (2004) Biochemistry, 43, pp. 16233-16242
  • Bertoncini, C.W., Jung, Y.S., Fernandez, C.O., Hoyer, W., Griesinger, C., Jovin, T.M., Zweckstetter, M., Release of long-range tertiary interactions potentiates aggregation of natively unstructured α-synuclein (2005) Proc. Natl Acad. Sci. USA, 102, pp. 1430-1435
  • Rochet, J.C., Conway, K.A., Lansbury Jr., P.T., Inhibition of fibrillization and accumulation of prefibrillar oligomers in mixtures of human and mouse α-synuclein (2000) Biochemistry, 39, pp. 10619-10626
  • Harper, J.D., Lansbury Jr., P.T., Models of amyloid seeding in Alzheimer's disease and scrapie: mechanistic truths and physiological consequences of the time-dependent solubility of amyloid proteins (1997) Annu. Rev. Biochem., 66, pp. 385-407
  • Selkoe, D.J., Folding proteins in fatal ways (2003) Nature, 426, pp. 900-904
  • Giese, A., Bader, B., Bieschke, J., Schaffar, G., Odoy, S., Kahle, P.J., Single particle detection and characterization of synuclein co-aggregation (2005) Biochem. Biophys. Res. Commun., 333, pp. 1202-1210
  • Masařík, M., Stobiecka, A., Kizek, R., Jelen, F., Pechan, Z., Hoyer, W., Sensitive electrochemical detection of native and aggregated α-synuclein protein involved in Parkinson's disease (2004) Electroanalysis, 16, pp. 1172-1181
  • Paleček, E., Ostatná, V., Masařík, M., Bertoncini, C.W., Jovin, T.M., Changes in interfacial properties of α-synuclein preceding its aggregation (2008) Analyst, 133, pp. 76-84
  • Hoyer, W., Antony, T., Cherny, D., Heim, G., Jovin, T.M., Subramaniam, V., Dependence of α-synuclein aggregate morphology on solution conditions (2002) J. Mol. Biol., 322, pp. 383-393
  • van Raaij, M.E., Segers-Nolten, I.M., Subramaniam, V., Quantitative morphological analysis reveals ultrastructural diversity of amyloid fibrils from α-synuclein mutants (2006) Biophys. J., 91, pp. L96-L98
  • Dusa, A., Kaylor, J., Edridge, S., Bodner, N., Hong, D.P., Fink, A.L., Characterization of oligomers during α-synuclein aggregation using intrinsic tryptophan fluorescence (2006) Biochemistry, 45, pp. 2752-2760
  • Kaylor, J., Bodner, N., Edridge, S., Yamin, G., Hong, D.P., Fink, A.L., Characterization of oligomeric intermediates in α-synuclein fibrillation: FRET studies of Y125W/Y133F/Y136F α-synuclein (2005) J. Mol. Biol., 353, pp. 357-372
  • Fernandez, C.O., Hoyer, W., Zweckstetter, M., Jares-Erijman, E.A., Subramaniam, V., Griesinger, C., Jovin, T.M., NMR of α-synuclein-polyamine complexes elucidates the mechanism and kinetics of induced aggregation (2004) EMBO J., 23, pp. 2039-2046
  • Wu, C.W., Yarbrough, L.R., N-(1-pyrene)maleimide: a fluorescent cross-linking reagent (1976) Biochemistry, 15, pp. 2863-2868
  • Karpovich, D.S., Blanchard, G.J., Relating the polarity-dependent fluorescence response of pyrene to vibronic coupling. Achieving a fundamental understanding of the py polarity scale (1995) J. Phys. Chem., 99, pp. 3951-3958
  • Dong, D.C., Winnik, M.A., The Py scale of solvent polarities (1984) Can. J. Chem., 62, pp. 2560-2565
  • Krishnan, R., Lindquist, S.L., Structural insights into a yeast prion illuminate nucleation and strain diversity (2005) Nature, 435, pp. 765-772
  • Lin, T.I., Excimer fluorescence of pyrene-tropomyosin adducts (1982) Biophys. Chem., 15, pp. 277-288
  • Tcherkasskaya, O., Davidson, E.A., Schmerr, M.J., Orser, C.S., Conformational biosensor for diagnosis of prion diseases (2005) Biotechnol. Lett., 27, pp. 671-675
  • Tamamizu-Kato, S., Kosaraju, M.G., Kato, H., Raussens, V., Ruysschaert, J.M., Narayanaswami, V., Calcium-triggered membrane interaction of the α-synuclein acidic tail (2006) Biochemistry, 45, pp. 10947-10956
  • Wahl, P., Analysis of fluorescence anisotropy decays by a least square method (1979) Biophys. Chem., 10, pp. 91-104
  • Celej, M. S., Jares-Erijman, E. A. & Jovin, T. M. (2008). Fluorescent N-arylaminonaphthalene sulfonate probes for amyloid aggregation of α-synuclein. Biophys. J. doi:10.1529/biophysj.107.125211; Hammarstrom, P., Kalman, B., Jonsson, B.H., Carlsson, U., Pyrene excimer fluorescence as a proximity probe for investigation of residual structure in the unfolded state of human carbonic anhydrase II (1997) FEBS Lett., 420, pp. 63-68
  • Winnik, F.M., Photophysics of preassociated pyrenes in aqueous polymer solutions and in other organized media (1993) Chem. Rev., 93, pp. 587-614
  • Piemonte, F., Caccuri, A.M., Morgenstern, R., Rosato, N., Federici, G., Aggregation of pyrene-labeled microsomal glutathione S-transferase. Effect of concentration (1993) Eur. J. Biochem., 217, pp. 661-663
  • Heise, H., Hoyer, W., Becker, S., Andronesi, O.C., Riedel, D., Baldus, M., Molecular-level secondary structure, polymorphism, and dynamics of full-length α-synuclein fibrils studied by solid-state NMR (2005) Proc. Natl Acad. Sci. USA, 102, pp. 15871-15876
  • Conway, K.A., Harper, J.D., Lansbury Jr., P.T., Fibrils formed in vitro from α-synuclein and two mutant forms linked to Parkinson's disease are typical amyloid (2000) Biochemistry, 39, pp. 2552-2563
  • Der-Sarkissian, A., Jao, C.C., Chen, J., Langen, R., Structural organization of α-synuclein fibrils studied by site-directed spin labeling (2003) J. Biol. Chem., 278, pp. 37530-37535
  • Serpell, L.C., Berriman, J., Jakes, R., Goedert, M., Crowther, R.A., Fiber diffraction of synthetic α-synuclein filaments shows amyloid-like cross-β conformation (2000) Proc. Natl Acad. Sci. USA, 97, pp. 4897-4902
  • Volles, M.J., Lansbury Jr., P.T., Relationships between the sequence of α-synuclein and its membrane affinity, fibrillization propensity, and yeast toxicity (2007) J. Mol. Biol., 366, pp. 1510-1522
  • Lashuel, H.A., Petre, B.M., Wall, J., Simon, M., Nowak, R.J., Walz, T., Lansbury Jr., P.T., α-synuclein, especially the Parkinson's disease-associated mutants, forms pore-like annular and tubular protofibrils (2002) J. Mol. Biol., 322, pp. 1089-1102
  • Uversky, V.N., Li, J., Fink, A.L., Evidence for a partially folded intermediate in α-synuclein fibril formation (2001) J. Biol. Chem., 276, pp. 10737-10744
  • Michl, J., Thulstrup, E.W., Spectroscopy with Polarized Light-Solute Alignment by Photoselection (1986) Liquid Crystals, Polymers and Membranes, p. 121. , VCH, UK
  • Uversky, V.N., A protein-chameleon: conformational plasticity of α-synuclein, a disordered protein involved in neurodegenerative disorders (2003) J. Biomol. Struct. Dynam., 21, pp. 211-234
  • Conway, K.A., Lee, S.J., Rochet, J.C., Ding, T.T., Williamson, R.E., Lansbury Jr., P.T., 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 (2000) Proc. Natl Acad. Sci. USA, 97, pp. 571-576
  • Allsop, D., Swanson, L., Moore, S., Davies, Y., York, A., El-Agnaf, O.M., Soutar, I., Fluorescence anisotropy: a method for early detection of Alzheimer β-peptide (Aβ) aggregation (2001) Biochem. Biophys. Res. Commun., 285, pp. 58-63
  • Weltman, J.K., Szaro, R.P., Frackelton Jr., A.R., Dowben, R.M., Bunting, J.R., Cathou, B.E., N-(3-pyrene)maleimide: a long lifetime fluorescent sulfhydryl reagent (1973) J. Biol. Chem., 248, pp. 3173-3177
  • Foguel, D., Suarez, M.C., Ferrao-Gonzales, A.D., Porto, T.C., Palmieri, L., Einsiedler, C.M., Dissociation of amyloid fibrils of α-synuclein and transthyretin by pressure reveals their reversible nature and the formation of water-excluded cavities (2003) Proc. Natl Acad. Sci. USA, 100, pp. 9831-9836
  • Ludescher, R.D., Peting, L., Hudson, S., Hudson, B., Time-resolved fluorescence anisotropy for systems with lifetime and dynamic heterogeneity (1987) Biophys. Chem., 28, pp. 59-75
  • Luk, K.C., Hyde, E.G., Trojanowski, J.Q., Lee, V.M., Sensitive fluorescence polarization technique for rapid screening of α-Synuclein oligomerization/fibrillization inhibitors (2007) Biochemistry, 46, pp. 12522-12529
  • Smith, W.S., Broadbridge, R., East, J.M., Lee, A.G., Sarcolipin uncouples hydrolysis of ATP from accumulation of Ca2+ by the Ca2+-ATPase of skeletal-muscle sarcoplasmic reticulum (2002) Biochem. J., 361, pp. 277-286

Citas:

---------- APA ----------
Thirunavukkuarasu, S., Jares-Erijman, E.A. & Jovin, T.M. (2008) . Multiparametric Fluorescence Detection of Early Stages in the Amyloid Protein Aggregation of Pyrene-labeled α-Synuclein. Journal of Molecular Biology, 378(5), 1064-1073.
http://dx.doi.org/10.1016/j.jmb.2008.03.034
---------- CHICAGO ----------
Thirunavukkuarasu, S., Jares-Erijman, E.A., Jovin, T.M. "Multiparametric Fluorescence Detection of Early Stages in the Amyloid Protein Aggregation of Pyrene-labeled α-Synuclein" . Journal of Molecular Biology 378, no. 5 (2008) : 1064-1073.
http://dx.doi.org/10.1016/j.jmb.2008.03.034
---------- MLA ----------
Thirunavukkuarasu, S., Jares-Erijman, E.A., Jovin, T.M. "Multiparametric Fluorescence Detection of Early Stages in the Amyloid Protein Aggregation of Pyrene-labeled α-Synuclein" . Journal of Molecular Biology, vol. 378, no. 5, 2008, pp. 1064-1073.
http://dx.doi.org/10.1016/j.jmb.2008.03.034
---------- VANCOUVER ----------
Thirunavukkuarasu, S., Jares-Erijman, E.A., Jovin, T.M. Multiparametric Fluorescence Detection of Early Stages in the Amyloid Protein Aggregation of Pyrene-labeled α-Synuclein. J. Mol. Biol. 2008;378(5):1064-1073.
http://dx.doi.org/10.1016/j.jmb.2008.03.034