Extended Data Fig. 9: Flowchart showing negative-stain electron microscopy data processing for human ACLY.
From: Structure of ATP citrate lyase and the origin of citrate synthase in the Krebs cycle
Starting from a final dataset of 27,293 particles, initial models were made using the stochastic gradient descent method in RELION2.1, applying C1, C2 or D2 symmetry. Subsequent 3D classification was performed using the C1, C2 or D2 starting models as an input, again applying C1, C2 or D2 symmetry, respectively. 3D classification using C1 and C2 symmetry clearly shows well-defined CCS modules in one-half of the hACLY molecule (C1: class 3, C2: class 3 and 4). Although 3D classification using D2 symmetry results in two classes displaying all four CCS modules (class 1 and 4), subsequent 3D refinement in C2 using an averaged map of these two classes resulted in a disappearance of two CCS modules in the lower half of hACLY, pointing to flexibility of the peripheral domains of hACLY.
