Fig. 2: Construction of an orthogonal mandi-regulated PYR1*MANDI/HAB1* dimerization module.
From: An orthogonalized PYR1-based CID module with reprogrammable ligand-binding specificity
a, Design pipeline for the development of PYR1*MANDI/HAB1*. b, Y2H assays of WT and orthogonal module components. c, PYR1*MANDI does not regulate WT HAB1 PP2C activity as measured using a 4-MUP substrate; data points indicate the mean of three technical replicates; and error bars indicate the s.d. The WT PYR1/HAB1 ABA response control in the left graph is the same as that used in Fig. 1c, as the datasets were acquired at the same time. d, The crystal structure of a PYR1*MANDI/mandi/HAB1* ternary complex demonstrates that the T162D/V393R orthogonalizing mutant pair installs a salt bridge. Coordinates for a PYR1-ABA-HAB1 complex (PDB 3QN1) were used to illustrate the WT interface. The images were rendered in Cinema3D using meshes exported from Pymol.
