Extended Data Fig. 2: Lipari–Szabo model-free analysis of 15N relaxation data of JIP1-SH3.
From: Visualizing protein breathing motions associated with aromatic ring flipping
a, Region of the 1H–15N HSQC spectrum of JIP1-SH3 at temperatures ranging from 5 to 46 °C. b, Experimental 15N R2 (CPMG) relaxation rates at a 1H frequency of 600 MHz and four different temperatures. c, Experimental 15N R1 relaxation rates at a 1H frequency of 600 MHz and four different temperatures. d, Experimental {1H}-15N heteronuclear NOEs acquired at a 1H frequency of 600 MHz and 25 °C. Error bars in b–d represent one standard deviation (s.d.) derived from Monte Carlo simulations of experimental uncertainty. e, A model-free analysis of 15N R1, R2 and heteronuclear NOEs at 25 °C was carried out providing an axially symmetric diffusion tensor (Supplementary Discussion). The diffusion tensor is represented relative to the dimeric structure of JIP1-SH3. Distributions of axis orientations are shown as red dots and were determined from Monte Carlo simulations using Tensor275. f, Angular dependence of the R2/R1 ratios relative to the main axis of the diffusion tensor of JIP1-SH3. Only residues without exchange contributions to the transverse relaxation and for which the {1H}-15N NOE is above 0.7 were included in the analysis. Error bars are centred at experimental values and were propagated from the experimental uncertainty on R2 and R1. Orange squares are back-calculated values using the optimal tensor. g, Order parameters, S2, derived from the model-free analysis of the relaxation data at 25 °C. Error bars represent one standard deviation (s.d.) derived from Monte Carlo simulations as implemented in Tensor2. h, Conformational exchange contributions, REX, derived from the model-free analysis of the relaxation data at 25 °C. Green bars indicate residues that are located in the dimer interface of the SH3 domain as detected by the PISA server76.
