Figure 5.

Characterization of the structure of DH-PH/RhoA complex in solution; (A) mapping of chemical shift perturbations within DH-PH upon binding to RhoA (magenta); two regions showing contacts involving RhoA and PH domain are shown in insets; residues exhibiting small chemical shift perturbations (Δσ < 0.05) are blue, medium (0.05 < Δσ < 0.10) are cyan and strongest (Δσ > 0.10) are green; residues that could be assigned only for free DH-PH and experience very strong perturbations in the complex are yellow; residues without assignment for free DH-PH are gray; chemical shift perturbations were calculated using the following formula sqrt(Δσ_HN²+0.1Δσ_N²). (B) An example of ¹H-¹⁵N TROSY-HSQC (red) and ¹H-¹⁵N HSQC (blue) for ²H,¹³C,¹⁵N-DH-PH/RhoA aligned in positively charged polyacrylamide gel, used to extract ¹D_HN RDCs; (C) fitting of RDCs measured for the DH-PH in the complex with RhoA to the crystal structure.