Extended Data Fig. 3: Structure and dynamics of the Y526A variant of JIP1-SH3.
From: Visualizing protein breathing motions associated with aromatic ring flipping
a, Comparison of the crystal structure of JIP1-SH3(Y526A) (green) with the WT structure (grey) showing an almost identical backbone conformation of the two proteins. The zoom highlights a minor structural difference at the level of the side chain of Q520 which reorients in the variant to take up the position normally occupied by Y526 in the WT protein. b, Superposition of 1H–15N HSQC spectra of JIP1-SH3(Y526A) (red) and JIP1-SH3(WT) (blue) acquired at 25 °C. c, Comparison of dihedral angles in JIP1-SH3(WT) (grey spheres) and JIP1-SH3(Y526A) (red lines). Dashed lines correspond to the backbone φ angle and full drawn lines to the backbone ψ angle. d, Conformational exchange contributions, REX, extracted from 15N CPMG relaxation dispersion data as the difference between R2eff at low (31 Hz) and high (1 kHz) CPMG frequencies. The exchange contributions are compared for WT JIP1-SH3 (grey, at 600 MHz) and the Y526A variant (red, at 700 MHz). e, Conformational exchange contributions, REX, extracted from 1HN CPMG relaxation dispersion data as the difference between R2eff at low (50 Hz) and high (2 kHz) CPMG frequencies. The exchange contributions are compared for the WT JIP1-SH3 (grey, at 600 MHz) and the Y526A variant (red, at 600 MHz). f–i, Chemical shift differences between the Y526A variant and WT JIP1-SH3 for 1HN (f), 15N (g), 13Cα (h) and 13Cβ (i).
