Change in pH during amyloid
formation of wild-type α-synuclein
and an α-synuclein 5Q mutant. (A) pH change during amyloid formation
detected by pH electrode. The proteins (wild-type α-synuclein
and the 5Q mutant) were isolated in pure water, and pH was measured
before and after fibril formation. On average, pH changed from pH
5.6 to pH 6.5 for the wild-type protein and on average from pH 6.4
to 6.6 for the 5Q mutants. (B) pH change detected using the pH sensitive
dye, resazurin. Monomeric α-synuclein in weak buffer containing
resazurin, at pH 5.8, resulted in purple color. The fibrillar sample
gave rise to blue color. For comparison, the pH was measured to be
6.4 using a pH electrode. (C) pH change detected using NMR spectroscopy
using the δ2 proton of histidine in monomeric α-synuclein
as a pH sensor. The red and blue traces show the spectrum before and
after fibril formation, respectively. The large decrease in chemical
shift detected for the monomer in the fibrillar sample is indicative
of a pH increase.