Extended Data Fig. 7: Analysis of CPMG relaxation dispersion data of the H493A variant of JIP1-SH3.
From: Visualizing protein breathing motions associated with aromatic ring flipping
a, Region of the 1H–15N HSQC of the H493A variant at three different temperatures (green - 15 °C, blue - 25 °C and red - 35 °C). b, Conformational exchange contributions, REX, extracted from 15N CPMG relaxation dispersion data of the H493A variant as the difference between R2eff at low (31 Hz) and high (1 kHz) CPMG frequencies (600 MHz and 15 °C). c, Conformational exchange contributions, REX, extracted from 1HN CPMG relaxation dispersion data of the H493A variant as the difference between R2eff at low (50 Hz) and high (2 kHz) CPMG frequencies (600 MHz and 15 °C). d, e, Examples of 15N (d) and 1HN (e) CPMG relaxation dispersion profiles of the H493A variant of JIP1-SH3 obtained at two magnetic field strengths (red – 600 MHz, blue – 950 MHz) at 15 °C. The 15N and 1HN data were analysed simultaneously for all residues according to a two-site exchange model (full-drawn lines in d and e) using a population of the minor state fixed to 10%. Error bars represent one standard deviation (s.d.) derived from Monte Carlo simulations of experimental uncertainty. f, g, Comparison of the chemical shift differences between the major and minor state extracted from relaxation dispersion experiments for WT JIP1-SH3 (red) and its H493A variant (blue). Data are shown for both 15N (f) and 1HN (g) chemical shifts.
