Amyloid fibrils are involved in several neuro-degenerative diseases. However, because of their polymorphism and low concentration, they are challenging to assess in real-time with conventional techniques. Here, we present a new approach for the characterization of the intermediates: protofibrils and "end-off" aggregates which are produced during the amyloid formation. To do so, we have fashioned conical track-etched nanopores that are functionalized to prevent the fouling. Using these nanopores, we have followed the kinetic of amyloid growth to discriminate the different intermediates protofibrils and "end-off. Then, the nanopore was used to characterize the effect of promoter and inhibitor of the fibrillation process. Finally, we have followed in real-time the degradation of amyloid with peptase. Compare with the SiN nanopore, the track-etched one features exceptionally high success rate via functionalization and detection in "one-pot". Our results demonstrate the potential for a conical nanopore to be used as a routine technique for the characterization of the amyloid growth and/or degradation.

Giamblanco, N., Coglitore, D., Gubbiotti, A., Ma, T., Balanzat, E., Janot, J.-., et al. (2018). Amyloid Growth, Inhibition, and Real-Time Enzymatic Degradation Revealed with Single Conical Nanopore. ANALYTICAL CHEMISTRY, 90(21), 12900-12908 [10.1021/acs.analchem.8b03523].

Amyloid Growth, Inhibition, and Real-Time Enzymatic Degradation Revealed with Single Conical Nanopore

Chinappi M.;
2018-01-01

Abstract

Amyloid fibrils are involved in several neuro-degenerative diseases. However, because of their polymorphism and low concentration, they are challenging to assess in real-time with conventional techniques. Here, we present a new approach for the characterization of the intermediates: protofibrils and "end-off" aggregates which are produced during the amyloid formation. To do so, we have fashioned conical track-etched nanopores that are functionalized to prevent the fouling. Using these nanopores, we have followed the kinetic of amyloid growth to discriminate the different intermediates protofibrils and "end-off. Then, the nanopore was used to characterize the effect of promoter and inhibitor of the fibrillation process. Finally, we have followed in real-time the degradation of amyloid with peptase. Compare with the SiN nanopore, the track-etched one features exceptionally high success rate via functionalization and detection in "one-pot". Our results demonstrate the potential for a conical nanopore to be used as a routine technique for the characterization of the amyloid growth and/or degradation.
2018
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore ING-IND/34 - BIOINGEGNERIA INDUSTRIALE
Settore ING-IND/06 - FLUIDODINAMICA
English
Amyloid; Curcumin; Dextran Sulfate; Ethanol; Hydrogen-Ion Concentration; Kinetics; Lactoglobulins; Pepsin A; Polyethylene Glycols; Quercetin; Nanopores; Protein Multimerization; Proteolysis
Giamblanco, N., Coglitore, D., Gubbiotti, A., Ma, T., Balanzat, E., Janot, J.-., et al. (2018). Amyloid Growth, Inhibition, and Real-Time Enzymatic Degradation Revealed with Single Conical Nanopore. ANALYTICAL CHEMISTRY, 90(21), 12900-12908 [10.1021/acs.analchem.8b03523].
Giamblanco, N; Coglitore, D; Gubbiotti, A; Ma, T; Balanzat, E; Janot, J-; Chinappi, M; Balme, S
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/247496
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