Figure 5.

The LCRs of Fip1₂₂₆ are highly dynamic within the CPF complex. (A) ¹H,¹⁵N 2D HSQC of Fip1₂₂₆, alone (red, left) and bound to CPFΔFip1 (blue, middle) or CPFΔFip1 and Pap1 (orange, right). Schematic diagrams show the proteins included in each experiment. All spectra were collected at 950 MHz with 11 µM ¹³C,¹⁵N,²H Fip1₂₂₆ in 150 mM NaCl buffer. Peaks analyzed in B are indicated in the spectra. BEST ¹H,¹⁵N-TROSY spectra were acquired with 64 scans and a recycle delay of 400 msec, giving a final spectral resolution of 2.2 Hz per point in the indirect dimension and an experimental time of 142 min. (B) Selected Fip1₂₂₆ peaks for all three samples. Perturbation or line broadening of peaks specific to the defined regions for Yth1 and Pap1 binding was observed upon interaction with CPFΔFip1 and Pap1. Peaks are mapped onto a diagram of the Fip1₂₂₆ protein. Colors as in A. (C) T2 relaxation data for the first 170 residues of Fip1₂₂₆ alone (red) and incorporated into CPFΔFip1 (blue).