Extended Data Fig. 1: Validation of biochemical activity, cryo-EM imaging, and image processing pipeline of the ApmFz2 ternary complex.
From: Structure of Fanzor2 reveals insights into the evolution of the TnpB superfamily
a. Biochemical validation of Fanzor2 (Fz2) cleavage activity tested on a linear DNA substrate containing a 5’-GGG TAM with either a correct target spacer (T) or a scrambled spacer (S) (n = 3). Red arrowheads indicate cleavage product. b. Representative cryo-EM micrograph from reconstituted ApmFz2 ternary complex (n = 3,205). Yellow circles indicate contaminates. Scale bar, 100 nm. c. 2D classification in cryoSPARC 4.4.0 on template-picked particles. Scale bar, 20 nm d. Image processing workflow used to analyze the cryo-EM data. e. Fourier shell correlation (FSC) curve. Resolution is reported at the 0.143 cut-off (solid blue line). f. Final ApmFz2 ternary complex refined reconstruction (right) and local resolution filtered reconstruction (left). Colors indicate the local resolution range, from 2.3 Å to 5.8 Å.
