Fig. 5: Binding kinetics suggest MAX and Pho4 differ in folding and binding transition. | Nature Communications

Fig. 5: Binding kinetics suggest MAX and Pho4 differ in folding and binding transition.

From: Mutations to transcription factor MAX allosterically increase DNA selectivity by altering folding and binding pathways

Fig. 5

a Example dissociation traces for WT (gray) and E32N (red) variants of MAX (top, n = 5 for all) and comparison of koff measurements (median ± SEM) across two devices and two DNA sequences (bottom). Dashed black line indicates 1:1 relationship. b Measured koff versus inferred kon for all Pho4 (orange) and MAX (teal) mutants interacting with cognate E-box (median ± SEM); dashed lines denote WT values. c Measured koff versus inferred kon for all Pho4 (top) and MAX (bottom) mutants interacting with cognate E-box (median ± SEM); dashed lines denote WT values. Marker color indicates mutations to known crystallographic DNA base contacts (red) or dimerization interface contacts (blue), also shown imposed on Pho4 (1A0A) and MAX (1HLO) crystal structures. WT and variant koffs were compared via independent t-test, and WT-like koff values (p ≥ 0.05) are indicated by light gray markers. d Measured koff and inferred kon (both median ± SEM) versus change in helical propensity39 for non-DNA contacting basic region substitutions in MAX (teal) and Pho4 (orange); dashed line indicates linear regression. Replicate counts for all Kd and koff measurements are contained in Supplementary Data 2. Source data are provided as a Source Data file.

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