Fig. 5: The tuning of the substrate-binding mode.
From: The evolutionary origin of naturally occurring intermolecular Diels-Alderases from Morus alba
a Overlay structures of ancDADS with moracin C (1) in blue and morachalcone A (4) in purple. b A Lineweaver–Burk plot showing competitive inhibition of the ancDADS-catalyzed oxidative cyclization reaction of moracin C (1) by dienophile 4. 100 μL enzyme reaction system: 1.5 μL of moracin C (20–150 μM), 0.3 μg of ancDADS, 50/150/500 μM of morachalcone A (4), 20 mM Tris-HCl buffer (pH 8.0). The data were presented as mean values ± standard error (SE), with error bars indicating the standard deviations of three independent measurements. c Frequency distribution histogram of the distance between FAD and the endo-transition state (TS). d The binding model of ancDADS-mut4-TS deduced from 500 ns MD simulations of the TS in complex with the enzyme. A black dashed line represents interactions between the substrate and surrounding residues. Nonconserved residues are indicated using red and blue fonts, while conserved residues are shown in black. e Evolution models of DAs from OCs.
