Supplementary Figure 2: Analysis of the DNCR2:danoprevir:NS3a complex crystal structure and the specificities of drug/NS3a complex reader proteins.
From: Multi-input chemical control of protein dimerization for programming graded cellular responses
a, 1 µM NS3a with avidity binding to yeast-displayed D5, DNCR1, or DNCR2. Technical triplicates and their mean are shown. b, Binding of 1 nM NS3a to DNCR2 displayed on the surface of yeast in the presence of increasing concentrations of danoprevir. Technical triplicates are shown. c, An overlay of DNCR2 (blue) from the DNCR2:danoprevir:NS3a complex with the original DHR79 scaffold (orange) crystal structure (PDBID: 5CWP) (Nature 528, 580-584, 2015) . Regions where there are modest changes in the backbone conformation are circled with a dotted line, including missing density for helix 8 and an unraveled helix 7 N-terminus. d, NS3a:danoprevir (blue) from the DNCR2:danoprevir:NS3a complex aligns closely to a crystal structure of NS3a:danoprevir (yellow) alone (PDBID: 3M5L) (PNAS 107, 20986-20991, 2010). e, Size exclusion chromatograms of DNCR2, NS3a, or DNCR2:NS3a complexes in the presence or absence of danoprevir. Representative of three technical replicates. f, Crystal structure of DNCR2:danoprevir:NS3a (blue) aligned to structures of asunaprevir:NS3a (lavender, PDBID: 4WF8) or grazoprevir:NS3a (yellow, PDBID: 3SUD) with clashes (red) between residues of DNCR2 and asunaprevir and grazoprevir highlighted (PLoS Pathog 8, e1002832, 2012; ACS Chem Biol 9, 2485-2490, 2014).
