Dynamics of the Pin1-CPEB1 pS210 complex and the direct involvement of the phosphate group in binding.
(a) Measurement of 1H, 15N NOE values for the apo Pin1 and (b) in complex with the pS210 ligand. (c) Local rotational correlation times τc were calculated from the T1/T2 ratio. Homogeneous values along the single domains were obtained: T1WW, apo = (612.7 ± 53.6) ms, T1Cat,apo = (792.7 ± 81.8) ms, T2WW,apo = (113.5 ± 6.9) ms and T2Cat,apo = (85.4 ± 13.1) ms; T1WW,pS210 = (767.1 ± 80.1) ms, T1Cat,,pS210 = (913.5 ± 91.3) ms, T2WW,pS210 = (94.1 ± 5.8) ms and T2Cat,pS210 = (74.8 ± 6.7) ms. (d) 1D- 31P NMR titration experiment of CPEB1pS210 with Pin1 WW domain. The black spectrum shows the free peptide; in blue the saturated complex with Pin1 WW is displayed. The phosphate buffer reference peaks hardly shows any chemical shift perturbation. (e) Chemical Shift Perturbations of the phosphate group of several CPEB1 peptides upon titration with Pin1 WW domain.