Fig. 3: Tryptophan-mediated quenching in lysozyme and MBP quenchbodies.
From: Optimised nanobody-based quenchbodies for enhanced protein detection
A Model of the lysozyme quenchbody (blue) highlighting all intrinsic tyrosines (yellow sticks) and tryptophans (red spheres, as labelled), in complex with lysozyme (grey surface model), based on PDB ID: 1ZVH. B Fold-sense for WT, W103Y, W115Y and W103Y/W115Y lysozyme quenchbody variants in the presence of 500 nM lysozyme. C Fold-sense for WT, Y27W, Y104W, Y110W, single-, double- and triple-mutant lysozyme quenchbody variants. D Model of the MBP quenchbody (blue) highlighting all intrinsic tyrosines (yellow sticks) and tryptophans (red spheres, as labelled), in complex with MBP (grey surface model), based on PDB ID: 5M14. E Fold-sense for WT, vs W101A/W110A/W115A triple mutant in the presence of 500 nM MBP. F Fold-sense for WT, Y33W, Y54W, Y59W and Y114W, double-, triple-, and quadruple-mutant MBP quenchbody variants. Data are mean ± SD (excitation = 535 ± 20 nm and emission = 585 ± 30 nm). Ordinary one-way ANOVA with Tukey’s multiple comparisons shows significant (**** = p < 0.0001) or non-significant (ns) differences.
