Fig. 3: Conformational states of the PAZ and Mid domain in the human Ago2 apo enzyme, binary and ternary complex.

Doubly labeled apo hAgo2^PAZ/Mid (A) and binary hAgo2^PAZ/Mid-RNA guide complexes (B) were immobilized via an hAgo2-directed antibody, whereas ternary hAgo2^PAZ/Mid-guide-target complexes (C) were immobilized via the target RNA^long-Biotin. FRET efficiency histograms show data from at least three independent measurements. All data were fitted using a Gaussian equation. D Fraction of molecules that show dynamics. Bars show the mean of at least three different measurements and error bars indicate the standard deviations (no. of biological replicates: apo enzyme = 4; binary complex = 3; ternary complex = 3). E Analysis of all dynamic molecules detected in the measurements presented in (A). Transitions between FRET states are presented in a transition density plot. F Comparison of FRET efficiency distributions of the apo hAgo2^PAZ/Mid enzyme and ternary complexes composed of hAgo2^PAZ/Mid, guide and target RNA. Source data provided as a Source Data file.