Extended Data Fig. 6: Conformational switching of ACLY during catalysis.

a, View of the interaction between the CCS and CCL modules in a representative hACLY-A/B crystal structure (space group P1), with the CCSα β-hairpin (orange) and CoA-binding domain (pink) in cartoon mode. Bound CoA is shown as coloured spheres. b, Overlay of a human CCL protomer in the closed state (pink CoA-binding domain) with the crystal structure of hACLY-A/B as in a (white CoA-binding domain). The resulting clash between the CoA-binding domain (with bound CoA-molecule) and the β-hairpin is indicated by a red box. c, View of the interaction between the CCS and CCL modules in the crystal structure of C. limicola ACLY-A/B, with the CCSα β-hairpin (orange) and CoA-binding domain (pink) in cartoon mode. d, Zoomed-in view of the stalk region in the crystal structures of hACLY-A/B and C. limicola ACLY-A/B based on the superposition of the helical core of the CCL modules. e, f, Interactions at the stalk region and β-hairpin as observed in the crystal structures of hACLY-A/B and C. limicola ACLY-A/B. g, Two-state rigid-body SAXS model for apo-hACLY-A/B (MultiFoXS, χ² = 2.8) overlaid with the hACLY-A/B crystal structures in space groups P1 and C2 (grey). h, Single-state rigid-body SAXS model for hACLY-A/B (MultiFoXS, χ² = 2.8) in the presence of both citrate and CoA overlaid with the hACLY-A/B crystal structures in space groups P1 and C2 (grey). i, Comparison between in-solution SAXS scattering profiles measured from linker-deleted hACLY-A/B and full-length hACLY. (i) Profiles recorded from hACLY-A/B (green) and hACLY (grey) in HBS buffer; (ii) profiles recorded from hACLY-A/B (purple) and hACLY (black) in HBS buffer supplemented with citrate and CoA; (iii) profiles recorded from hACLY in HBS buffer (grey) and HBS buffer supplemented with both citrate and CoA (black); (iv) profiles recorded from hACLY-A/B in HBS buffer (green) and HBS buffer supplemented with both citrate and CoA (purple); and (v) fit of the theoretical scattering profile (red) calculated from an AllosMod-FoXS model for hACLY (as shown in j) to the experimental scattering profile recorded in the presence of citrate and CoA (black). j, AllosMod-FoXS SAXS model for hACLY in HBS buffer supplemented with citrate and CoA, overlaid with the hACLY-A/B crystal structures in space groups P1 and C2 (grey). In g, h and j, the bottom numeric table presents an all-residue (Cα) r.m.s.d. matrix for the hACLY-A/B crystal structures and presented SAXS models, and for each crystal structure and model the calculated fit (χ² value) against the recorded SAXS data are shown as calculated by FoXS, Crysol and Crysol 3.0. P1-hACLY-A/B_1 and P1-hACLY-A/B_2 denote structures for hACLY-A/B extracted from the P1 crystal form; C2-hACLY-A/B_1 and C2-hACLY-A/B_2 denote structures for hACLY-A/B extracted from the C2 crystal form.