Fig. 4.

Diverse Rubisco enzymes vary in their tendency to demix with EPYC1. a, b The tendency of a phylogenetically diverse set of Rubisco enzymes to demix was assessed using the sedimentation assay (a) and microscopy (b). Scale bar 15 µm. c, d The role of the small subunit in demixing was assessed using chimeric Rubiscos (Synechococcus large subunit cores (L₈) either lacking, or assembled with small subunits from rice (Os) or Chlamydomonas (Cr). Sedimentation (c) and microscopy (d) assays are shown. Scale bar 15 µm. e Addition of external CrRbcS (4 µM) rapidly changes the appearance of SeL₈ (2.5 µM)-EPYC1/EPYC1-GFP (1.25/0.1 µM) droplets. Scale bar 5 µm. f Native PAGE gel-shift assay using Rubisco and the single-repeat variant EPYC1Δ2-4 indicates increased binding for Rubiscos lacking the small subunit