Fig. 2: ¹H-¹⁵N spectra of WT, D121A and pS129 aSyn indicate conformational differences.

a We compared chemical shift perturbations in the amide backbone of ¹H-¹⁵N D121A aSyn to WT aSyn (green) and pS129 aSyn to WT aSyn (yellow) in 20 mM Tris pH 7.2 (using a fixed protein concentration of 200 μM) and observed CSPs around the location of the D to A mutation and the PO₄²⁻ of S129 at the C-terminus (for the full zoomed out spectrum see Supplementary Fig. 3.). (b) Significant CSPs were observed at the C-terminus upon addition of 4.2 mM calcium for all, WT (blue), D121A (green) and pS129 (yellow) aSyn. (Titration data for all concentrations of calcium are available in Supplementary Fig. 4). Higher CSPs are observed for D121A compared to WT and pS129 aSyn. c ¹H-¹⁵N HSQC NMR spectra of pS129 and D121A aSyn in the absence (red) and in the presence of calcium (green) (Supplementary Figs. 6 and 7 show spectra with more labels). Major CSPs in the presence of calcium are located at the C-terminus (arrows with assigned amino acid residues) in both pS129 and D121A aSyn.