Extended Data Fig. 2: Glp-FtsZ interaction.
(a) BLI sensorgrams of FtsZ binding to immobilized Glp in the presence or absence of 1 mM GTP. (b) Normalized melting curves of Glp with or without 1 mM FtsZCTD peptide as determined by a thermofluor assay. Glp was stabilized by 2 °C in the presence of FtsZCTD. (c) The crystal structures of ligand-free (red) and FtsZCTD-bound (blue and yellow ligand). Glp can be superimposed with an r.m.s.d. of 0.64 Å for 392 equivalent Cα atoms. (d) Electron density map of FtsZCTD bound to Glp monomer B contoured at 1.2 σ. (e) Comparison of the Glp-FtsZCTD complex (red, left panel) with the Gephyrin-GlyR complex (blue, right panel, PDB code: 2fts). In both cases the FtsZCTD and GlyR peptides (molecular surfaces) bind the domain IV (in colour) of Glp and gephyrin respectively. (f) Far-UV circular dichroism spectra of Glp and GlpΔloop. (g) BLI sensorgrams of FtsZ (10 μM) binding to immobilized Glp or GlpΔloop. (h) Representative images for mNeon-GlpΔloop in Cglu_Δglp. Scale bar = 5 μm. (i) Left, Violin plots showing the distribution of cell length. The number of cells used in the analyses (n) is indicated below each plot representing triplicate experiments. The box indicates the 25th to the 75th percentile, the mean and the median are indicated with a dot and a line in the box, respectively. Significance indicated corresponds to values of Cohen’s d (from top to bottom: (***, d = 1,55, P = 0), (ns, d = 0,09, P = 0,0012), (***, d = 1,76, P = 0), (***, d = 1,58, P = 0), (***, d = 1,78, P = 0), (ns, d = 0, P = 0,95)). Right, frequency histogram indicating the number of septa per cell, calculated from n cells imaged from 3 independent experiments (triplicates) for each strain (for Cglu, n = 718, 1468 and 1223; for Cglu_Δglp + mNeon-Glp, n = 1641, 1311 and 1801; for Cglu_Δglp, n = 873, 1538 and 840; for Cglu_Δglp + mNeon-GlpΔloop, n = 678, 1187 and 905); open circles represent the corresponding data points; bars represent the mean ± SD. Cohen’s d from top to bottom (ns, d = 0.28, P = 1.19e-26), (***, d = 1.25, P = 0).
