Sensitive Electrochemical Detection of Native and Aggregated x-Synuclein Protein Involved in Parkinson's Disease

Michal Masarik, Agata Stobiecka, René Kizek, Frantisek Jelen, Zdenk Pechan, Wolfgang Hoyer, Vinod Subramaniam, Emil Palecek

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Abstract

The aggregation of α-synuclein, a 14 kDa protein, is involved in several human neurodegenerative disorders, including Parkinson's disease. We studied native and in vitro aggregated α-synuclein by circular dichroism (CD), atomic force microscopy (AFM) and electrochemical methods. We used constant current chronopotentiometric stripping analysis (CPSA) to measure hydrogen evolution catalyzed by α-synuclein (peak H) at hanging mercury drop electrodes (HMDE) and square-wave stripping voltammetry (SWSV) to monitor tyrosine oxidation at carbon paste electrodes (CPE). To decrease the volume of the analyte, most of the electrochemical measurements were performed by adsorptive transfer (medium exchange) from 3-6 L drops of α-synuclein samples. With both CPE and HMDE we observed changes in electrochemical responses of α-synuclein corresponding to protein fibrillization detectable by CD, fluorescence and AFM. Aggregation-induced changes in peak H at HMDE were relatively large in strongly aggregated samples, suggesting that this electrochemical signal may find use in the analysis of early stages of α-synuclein aggregation. This assumption was documented by marked changes in the peak H potential and height in samples withdrawn at the end of the lag and the beginning of the elongation phase. Native α-synuclein can be detected down to subnanomolar concentrations by CPSA.
Original languageUndefined
Pages (from-to)1172-1181
Number of pages10
JournalElectroanalysis
Volume16
Issue number13-14
DOIs
Publication statusPublished - 2004

Keywords

  • METIS-217999
  • α-synuclein aggregation
  • Electrochemistry of proteins
  • Mercury and carbon electrodes
  • Adsorptive transfer stripping
  • IR-71994
  • Catalytic hydrogen evolution

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