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Abstract:

α-synuclein is a major component of intraneuronal protein aggregates constituting a distinctive feature of Parkinson disease. To date, fluorescence imaging of dynamic processes leading to such amyloid deposits in living cells has not been feasible. To address this need, we generated a recombinant α-synuclein (α-synuclein-C4) bearing a tetracysteine target for fluorogenic biarsenical compounds. The biophysical, biochemical and aggregation properties of α-synuclein-C4 matched those of the wild-type protein in vitro and in living cells. We observed aggregation of α-synuclein-C4 transfected or microinjected into cells, particularly under oxidative stress conditions. Fluorescence resonance energy transfer (FRET) between FlAsH and ReAsH confirmed the close association of fibrillized α-synuclein-C4 molecules. α-synuclein-C4 offers the means for directly probing amyloid formation and interactions of α-synuclein with other proteins in living cells, the response to cellular stress and screening drugs for Parkinson disease.

Registro:

Documento: Artículo
Título:Fluorescence imaging of amyloid formation in living cells by a functional, tetracysteine-tagged α-synuclein
Autor:Roberti, M.J.; Bertoncini, C.W.; Klement, R.; Jares-Erijman, E.A.; Jovin, T.M.
Filiación:Department of Molecular Biology, 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:alpha synuclein; amyloid; amyloid; cysteine; fluorescent dye; hybrid protein; reactive oxygen metabolite; article; cell stress; fluorescence analysis; fluorescence resonance energy transfer; oxidative stress; Parkinson disease; priority journal; protein aggregation; protein protein interaction; chemistry; confocal microscopy; Escherichia coli; fluorescence microscopy; fluorescence resonance energy transfer; gene vector; genetic procedures; genetic transfection; genetics; human; metabolism; methodology; tumor cell line; alpha-Synuclein; Amyloid; Biosensing Techniques; Cell Line, Tumor; Cysteine; Escherichia coli; Fluorescence Resonance Energy Transfer; Fluorescent Dyes; Genetic Vectors; Humans; Microscopy, Confocal; Microscopy, Fluorescence; Oxidative Stress; Reactive Oxygen Species; Recombinant Fusion Proteins; Transfection
Año:2007
Volumen:4
Número:4
Página de inicio:345
Página de fin:351
DOI: http://dx.doi.org/10.1038/nmeth1026
Título revista:Nature Methods
Título revista abreviado:Nat. Methods
ISSN:15487091
CAS:alpha synuclein, 154040-18-3; amyloid, 11061-24-8; cysteine, 4371-52-2, 52-89-1, 52-90-4; alpha-Synuclein; Amyloid; Cysteine, 52-90-4; Fluorescent Dyes; Reactive Oxygen Species; Recombinant Fusion Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15487091_v4_n4_p345_Roberti

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Citas:

---------- APA ----------
Roberti, M.J., Bertoncini, C.W., Klement, R., Jares-Erijman, E.A. & Jovin, T.M. (2007) . Fluorescence imaging of amyloid formation in living cells by a functional, tetracysteine-tagged α-synuclein. Nature Methods, 4(4), 345-351.
http://dx.doi.org/10.1038/nmeth1026
---------- CHICAGO ----------
Roberti, M.J., Bertoncini, C.W., Klement, R., Jares-Erijman, E.A., Jovin, T.M. "Fluorescence imaging of amyloid formation in living cells by a functional, tetracysteine-tagged α-synuclein" . Nature Methods 4, no. 4 (2007) : 345-351.
http://dx.doi.org/10.1038/nmeth1026
---------- MLA ----------
Roberti, M.J., Bertoncini, C.W., Klement, R., Jares-Erijman, E.A., Jovin, T.M. "Fluorescence imaging of amyloid formation in living cells by a functional, tetracysteine-tagged α-synuclein" . Nature Methods, vol. 4, no. 4, 2007, pp. 345-351.
http://dx.doi.org/10.1038/nmeth1026
---------- VANCOUVER ----------
Roberti, M.J., Bertoncini, C.W., Klement, R., Jares-Erijman, E.A., Jovin, T.M. Fluorescence imaging of amyloid formation in living cells by a functional, tetracysteine-tagged α-synuclein. Nat. Methods. 2007;4(4):345-351.
http://dx.doi.org/10.1038/nmeth1026