Fig. 1: The identification and characterisation of cyclic peptide (CP) inhibitors of ADO.
a A schematic highlighting the key steps of RaPID. Generated using Adobe Illustrator, Revvity Chemdraw and CCP4MG. Protein coordinates were obtained from the PDB using accession code 8UAN. b A sequence alignment of the top eight unique CPs discovered for ADO. c Single-dose inhibition assays: Relative enzymatic activity of ADO with RGS52-15 (100 µM) in the presence of CP (10 µM) at 37 °C. The average of three independent experiments (n = 3) is shown (error bars show the standard error). d Biophysical analysis and inhibitory responses of CP1 and CP6. (Left) Single-cycle kinetic (SCK) SPR sensorgrams of CP1 and CP6 with ADO. The sensorgrams are shown in red and the fits to the data are shown in black. The concentrations of CPs used in the titration and the equilibrium dissociation constants (KD) are shown (KD given as the geometric mean of a minimum of three independent SPR measurements (n = 3) with standard error). (Middle) Dose-response curves with the half-maximal inhibitory concentration (IC50) values for CP1 and CP6. The average of three independent experiments (n = 3) is shown (error bars show the standard error). (Right) Michaelis-Menten kinetic plots of ADO activity in the absence and presence of CP1 and CP6 performed under aerobic conditions at 37 °C. The average of three independent experiments (n = 3) is shown (error bars show the standard error). Source data are provided as Source Data file.
