A FRET Sensor for Non-Invasive Imaging of Amyloid Formation in Vivo

Autor: Ellen A. A. Nollen, Jeremy N. Skepper, Gabriele S. Kaminski Schierle, Carlos W. Bertoncini, Simon Schlachter, Stefanie Schwedler, Tjakko J. van Ham, Alessandro Esposito, Janet R. Kumita, Clemens F. Kaminski, Fiona T. S. Chan, Annemieke T. van der Goot, Christopher M. Dobson
Přispěvatelé: University of Groningen, Molecular Neuroscience and Ageing Research (MOLAR)
Jazyk: angličtina
Rok vydání: 2011
Předmět:
FLIM
Amyloid
Recombinant Fusion Proteins
Protein aggregation
010402 general chemistry
01 natural sciences
TOXICITY
Article
BETA PLAQUES
03 medical and health sciences
chemistry.chemical_compound
RED-BLOOD-CELLS
synuclein
PARKINSONS-DISEASE
Bacterial Proteins
In vivo
protein folding
Cell Line
Tumor
Fluorescence Resonance Energy Transfer
Animals
Humans
Physical and Theoretical Chemistry
FLUORESCENCE
Caenorhabditis elegans
STATE NMR-SPECTROSCOPY
030304 developmental biology
Luminescent Proteins
Alpha-synuclein
0303 health sciences
POLYGLUTAMINE AGGREGATION
Chemistry
amyloid beta-peptides
biosensors
ALPHA-SYNUCLEIN AGGREGATION
Atomic and Molecular Physics
and Optics
0104 chemical sciences
Kinetics
Förster resonance energy transfer
Biochemistry
Microscopy
Fluorescence
MISFOLDING DISEASES
Synuclein
Biophysics
Protein folding
PROTEIN AGGREGATION
Zdroj: Chemphyschem, 12(3), 673-680. WILEY-V C H VERLAG GMBH
ISSN: 1439-4235
Popis: Misfolding and aggregation of amyloidogenic polypeptides lie at the root of many neurodegenerative diseases. Whilst protein aggregation can be readily studied in vitro by established biophysical techniques, direct observation of the nature and kinetics of aggregation processes taking place in vivo is much more challenging. We describe here, however, a Forster resonance energy transfer sensor that permits the aggregation kinetics of amyloidogenic proteins to be quantified in living systems by exploiting our observation that amyloid assemblies can act as energy acceptors for variants of fluorescent proteins. The observed lifetime reduction can be attributed to fluorescence energy transfer to intrinsic energy states associated with the growing amyloid species. Indeed, for alpha-synuclein, a protein whose aggregation is linked to Parkinson's disease, we have used this sensor to follow the kinetics of the self-association reactions taking place in vitro and in vivo and to reveal the nature of the ensuing aggregated species. Experiments were conducted in vitro, in cells in culture and in living Caenorhabditis elegans. For the latter the readout correlates directly with the appearance of a toxic phenotype. The ability to measure the appearance and development of pathogenic amyloid species in a living animal and the ability to relate such data to similar processes observed in vitro provides a powerful new tool in the study of the pathology of the family of misfolding disorders. Our study confirms the importance of the molecular environment in which aggregation reactions take place, highlighting similarities as well as differences between the processes occurring in vitro and in vivo, and their significance for defining the molecular physiology of the diseases with which they are associated.
Databáze: OpenAIRE
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