Extended Data Fig. 1: Structural propensities and conformational exchange in JIP1-SH3.
From: Visualizing protein breathing motions associated with aromatic ring flipping
a, 1H–15N HSQC spectrum of JIP1-SH3 at 25 °C with labels indicating assignments. Horizontal lines connect side chain resonances. b, Secondary structure propensities of JIP1-SH3 calculated from experimental 13Cα and 13Cβ chemical shifts at 25 °C. The position of secondary structure elements is indicated, as observed in the crystal structure of JIP1-SH3 (PDB: 2FPE). c, Differential scanning fluorimetry (DSF) melting curve for JIP1-SH3 (RFU – relative fluorescence unit). d, Conformational exchange contributions, REX, extracted from 15N CPMG relaxation dispersion data acquired at 15 °C of JIP1-SH3 (blue – 850 MHz, red – 600 MHz) and JIP1-SH3(Y526A) (orange – 700 MHz) as the difference between R2eff at low (31 Hz) and high (1 kHz) CPMG frequencies. e, Exchange contributions, REX, extracted from 1HN CPMG relaxation dispersion data acquired at 15 °C of JIP1-SH3 (blue – 950 MHz, red – 600 MHz) and JIP1-SH3(Y526A) (orange – 600 MHz) as the difference between R2eff at low (50 Hz) and high (2 kHz) CPMG frequencies. f, Structure of the dimeric SH3 domain of JIP1 with Y526 shown in cyan. Residues with exchange contributions (15N REX > 3 s−1 and 1HN REX > 10 s−1) are shown as spheres with colours indicating their position in the primary sequence (region 1 – green, region 2 – red, region 3 – blue). The residues showing 15N and 1HN exchange contributions are represented separately on each monomer.
