Figure 7.

Proximal-to-distal allostery confirmed by the mutant R95E_R352E. Titration of Adx interferes with cYY binding by spectral binding assay (A), likely by promoting a closed conformation of the enzyme. The significance of Adx0 and Adx40 for both datasets was extracted by t test (p < 0.05). The double proximal mutant R95E_R352E was designed to prevent the CYP-redox partner complex and did not show catalytic activity (B) or the ability to cross-link with Adx (C). The double CYP121 mutant was also combined with S171C to ¹⁹F label and monitor effects at the FG-loop (D) and (E).