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. 2017 Oct 13;429(20):3018–3030. doi: 10.1016/j.jmb.2017.09.005

Fig. 1.

Fig. 1.

DiY formation kinetics of WT α-syn and Y-to-A mutants. (a) DiY formation kinetics of WT α-syn and single/double/triple Y-to-A mutants upon UV irradiation at 280 nm wavelength, monitored by DiY fluorescence at 410 nm. Fluorescence intensities were normalized by setting the highest fluorescence value of WT DiY-α-syn to unity. All curves are means of triplicate measurements. Solid lines represent fits to a consecutive two-step reaction, except for WT α-syn, for which a one-step first-order model was applied. (b) Zoom into the first 10 s of the kinetics of WT α-syn and single Y-to-A mutants. (c) Zoom into the DiY formation kinetics of the triple Y-to-A mutant, Y125A–Y133A–Y136A. (d) α-Syn variants investigated in this study. (e) SDS-PAGE of WT α-syn and Y-to-A mutants before (−) and after (+, samples taken when the DiY fluorescence plateau was reached) UV irradiation. 1: WT α-syn, 2–9: single/double/triple Y-to-A mutants in same order as in panel d, 10: quadruple Y-to-A tyrosine knockout mutant. (f) SEC of WT DiY-α-syn, C-term DiY-α-syn (Y39A), and N-term DiY-α-syn (Y125A–Y133A–Y136A). (g) DiY fluorescence in dimer and monomer peaks after SEC of WT DiY-α-syn, total fluorescence, and fluorescence intensity normalized to protein concentration are shown as means of triplicates.