Early oligomeric aggregates of human alpha-synuclein are implicated in interactions with essential cellular components leading to toxicity. Very little is known about the molecular details of these aggregate species. We have developed a method that uses sub-stoichiometric labeling, that is, only a fraction of the monomers contain a fluorescent label, in combination with single-molecule photobleaching to determine the number of monomers per oligomer (Zijlstra et al., Angew Chem Int Ed Engl 51, 8821–8824, 2012). The number of bleaching steps gives the number of fluorescent labels per oligomer. Knowing the exact label density, that is, the fraction of labeled monomers at the start of the aggregation, we can correlate the number of fluorescent labels per oligomer to the total number of monomers. Using this method, we can determine the composition, probe the distribution in the number of monomers per oligomer, and investigate the influence of the fluorescent label on the aggregation process.
|European biophysics journal
|Published - 2013
|9th European Biophysics Congress, EBSA 2013 - Lisbon, Portugal
Duration: 13 Jul 2013 → 17 Jul 2013
Conference number: 9