Titration of RTshort-
(A) Overlay of TROSY-1H/15N-HSQC spectra of 205 μM RTshort-mt4 recorded at a sample temperature of 288 K during the titration with different ATP/protein ratios. Key: black: 0; red: 15; green: 48; cyan: 92; blue: 142; pink: 195. Blow-ups of the framed areas (a) and (b) are shown underneath the spectra and represent examples of significant chemical shift changes with RTshort-mt4 and ATP. (B) Determination of the mean KD-value for ATP with RTshort-mt4. Seven fits for normalized chemical shift changes of seven shifting residues encircled in (A) were used to calculate the mean KD-value with standard deviation. Exemplified, fitted curves for three signal changes depicted in boxes (a) and (b) are shown as a function of the [ATP]/[RTshort-mt4] ratio.