Figure 5
From: A new regulatory mechanism for Raf kinase activation, retinoic acid-bound Crabp1
NMR analyses showing Crabp1 direct interaction with the RBD of cRaf. (A) An expanded region of 1H,15N HSQC spectra of the 19 µM Crabp1 in the absence (black peaks) and presence (red peaks) of 126 µM cRaf-(R) (aa 51–187). (B) Titration curves of selected Crabp1 residues (C96, T97, Q98) in which the chemical shift change is plotted vs. the cRaf-(R) concentration. (C) Maximal chemical shift changes (∆δ) induced by cRaf-(R) on Crabp1. (D) Maximal resonance intensity changes (∆Intensity) induced by cRaf-(R) on Crabp1. (E) Cartoon representation of cRaf-(R) interaction sites with marked spectral changes on Crabp1 in the presence of cRaf-(R): 1 SD – orange, 2 SD – red. PDB: 1CBR. Chemical shifts were internally referenced to DSS (4,4-dimethyl-4-silapentane-1-sulfonic acid), and chemical shift differences in panels (C,D) were calculated as [(∆δ 1H)2 + (0.25 ∆δ 15N)2]1/2. NMR acquisition and sample conditions are described in the “Material and Methods”.
