Fig. 3. NMR analysis of P1 and C2 binding to ASF1.

(A) Overlay of the SOFAST HMQC spectra of ¹⁵N-hASF1A alone and after addition of a twofold excess of P1 or C2 (in gray, green, and magenta, respectively). The region indicated with a black box is magnified in (B, left and middle). (B) Top and middle: Overlay of five spectra measured at different concentrations of P1 and C2, respectively, with a rainbow color code from blue to red (in dark blue, free hASF1A, and in red, after addition of twofold excess of peptide/chimera), for the region boxed in (A) showing the modification signals of Thr⁹³ and Ile⁹⁷ upon titration. The position of the signal in the bound state is indicated by an arrow. Bottom: Representative titration curve for ASF1 residues Thr⁹³ and Ile⁹⁷ using the same color code. The fitted curve is depicted as a dashed line. (C) Chemical shift mapping on hASF1A surface after addition of a twofold excess of P1 or C2 as indicated. Increasing color scale corresponds to higher chemical shift variations as in fig. S4.