Extended Data Fig. 9: The A541L variant does not capture the structural details of the minor state detected by CPMG relaxation dispersion in JIP1-SH3.
From: Visualizing protein breathing motions associated with aromatic ring flipping
a, Superposition of 1H–15N HSQC spectra of JIP1-SH3(A541L) (red) and WT JIP1-SH3 (blue) acquired at 15 °C. b, Conformational exchange contributions, REX, extracted from 15N CPMG relaxation dispersion data of the A541L variant as the difference between R2eff at low (31 Hz) and high (1 kHz) CPMG frequencies at 700 MHz and 15 °C. Only modest 15N conformational exchange contributions are observed suggesting that this variant is populating a single conformation in solution represented by the determined crystal structure. c, d, Comparison of the chemical shift differences between the major and minor states extracted from relaxation dispersion experiments of WT JIP1-SH3 (blue) and the chemical shift differences between the observed chemical shifts of the A541L variant and WT JIP1-SH3 (red). Data are shown for 15N (c) and for 1HN (d) nuclei. The poor agreement between the two datasets show that the A541L crystal structure is not representative of the conformation of the minor state detected by NMR relaxation dispersion. e, f, Chemical shift differences between the A541L variant and WT JIP1-SH3 for 13Cα (e) and 13Cβ (f).
