Fig. 5: Competitive binding of Dab2_320–495 and Eps15_IDR to EH123.

A CSPs calculated between the ¹H–¹⁵N TROSY-HSQC of 200 µM ¹⁵N EH123 in the absence and presence of 200 µM Dab2_320–495 and/or 200 µM Eps15_IDR. Below are ¹⁵N R_1ρ spin relaxation rates of ¹⁵N EH123 in the absence and presence of 100% Eps15_IDR and both 100% Dab2_320–495 and Eps15_IDR at a ¹H frequency of 1200 MHz. Only the R_1ρ rates within EH1 in the presence of Eps15_IDR and Eps15_IDR with Dab2_320–495 are shown as the peaks of EH2 and EH3 were severely broadened. The parts of EH123 corresponding to EH1, EH2, and EH3 are illustrated above the plots. B CSPs calculated between the ¹H-¹⁵N HSQC of 100 µM ¹⁵N Eps15_IDR in the absence and presence of 100 µM EH123 and 100 µM EH123 + 100 µM Dab2_320–495, and ¹⁵N R_1ρ spin relaxation of ¹⁵N Eps15_IDR in the absence and presence of 100% EH123 and 100% EH123 + 100% Dab2_320–495 at a ¹H frequency of 1200 MHz. C ¹H–¹⁵N HSQC spectra of 100 µM ¹⁵N Eps15_IDR alone and in the presence of 100 µM EH123. D Zoom into ¹H-¹⁵N HSQC spectrum of 100 µM ¹⁵N Eps15_IDR alone and in the presence of 100 µM EH123 and 100 µM EH123 + 100 µM Dab2_320–495. E Zoom into ¹H-¹⁵N HSQC spectrum of 100 µM ¹⁵N Dab2_320–495 alone and in the presence of 100 µM EH123 and 100 µM EH123 + 100 µM Eps15_IDR. Color codes are denoted in the respective panels. The relaxation rates in (A) and (B) (lower panels) were derived from a fit of peak intensities against the relaxation delay. Errors of the fitted rates were derived from the experimental uncertainty.