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. Author manuscript; available in PMC: 2022 Sep 2.
Published in final edited form as: Structure. 2021 May 19;29(9):1048–1064.e6. doi: 10.1016/j.str.2021.05.002

Figure 5. Slow conformational transitions of α-syn observed by smPIFE experiments.

Figure 5.

(A) The histograms of average fluorescence lifetimes from smPIFE measurements of sCy3 labelled α-syn. sCy3 dye is conjugated at Cys in the V26C, Y39C or A56C mutants, respectively. The ensemble of α-syn includes different subpopulations with various degrees of dye available volume that interconvert slower than a few milliseconds (slower than burst durations).

(B) and (C) The histograms of average fluorescence lifetimes when sCy3 is labeling bases of double or single stranded DNA, respectively. The labeling positions were the promoter registers in the template strand of the lacCONS promoter (Kim et al., 2011; Mukhopadhyay et al., 2001; Ploetz et al., 2016). Note the low mean lifetime values in flexible single stranded DNA and the slightly higher mean values in more rigid and more sterically restricted double stranded DNA.