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. 2022 Oct 6;13:5892. doi: 10.1038/s41467-022-33552-x

Fig. 2. Cooperative binding of DND1’s tandem RRMs to AU-rich RNA.

Fig. 2

A ITC measurement of DND1’s tandem RRM domains titrated with CUUAUUUG RNA. N = 1.04 +/− 0.004; KD = 0.69 +/− 0.03 uM; ΔH = −31.4 +/− 0.2 kcal/mol; −TΔS = 23.0 kcal/mol. B overlay of two 1H-15N HSQC spectra of the free DND1 tandem RRMs (in blue) and a 1:1 complex with CUUAUUUG RNA (in red) demonstrates cooperative RNA binding. NMR conditions: Protein concentration 0.2 mM, 298 K, 750 MHz. NMR/ITC buffer: 100 mM Potassium Phosphate pH 6.6, 1 mM DTT. C ITC measurement of DND1’s extended RRM1 titrated with CUUAUUUG (N = 1 (set); KD > 54 μM; ΔH = −35 +/− 2 kcal/mol; −TΔS = 29.4 kcal/mol). D ITC measurement of DND1’s tandem RRMs titrated with UUUUUAC (N = 1 (set); KD > 35 μM; ΔH = −30.1 +/− 0.7 kcal/mol; −TΔS = 24.5 kcal/mol) and E DND1’s tandem RRM domains titrated with UUUUUCC (N = 1 (set); KD > 100 μM; ΔH = −41 +/− 4 kcal/mol; −TΔS = 35 kcal/mol). See also Supplementary Fig. 3 and Supplementary Table 2.