Extended Data Fig. 3: PspA rod radial density profiles and mass per length.
From: Structural plasticity of bacterial ESCRT-III protein PspA in higher-order assemblies
A: Cryo-EM density fit of atomic PspA models of different diameters focusing on the helices α1/α2 hairpin and helix 5 (α1 red, α2 + 3 violet, α4 blue, α5 cyan). (180 Å: PDB 8AKQ, 200 Å: PDB 8AKR, 215 Å: PDB 8AKS, 235 Å: PDB 8AKT, 250 Å: PDB 8AKU, 270 Å: PDB 8AKV, 280 Å: PDB 8AKW, 290 Å: PDB 8AKY, 305 Å: PDB 8AKX, 320 Å: PDB 8AKZ, 365 Å: PDB 8AL0) B: Scatter plot of three selected pairs (R44-L185, E126-S163, E179-K55) distance changes with respect to the initial distance in the 180 Å diameter assembly over all rod diameters. C: Box plot of pair distance changes of evolutionarily conserved residues with respect to the initial distance in the 180 Å diameter assembly. The residues were selected by first identifying potential intermolecular interactions between highly conserved residues ( > 90% conserved among PspA/Vipp1 proteins9. Then, the Cα distance for each pair was measured for each rod diameter. To calculate the difference of the pair distances relative to the smallest diameter rods, the distances in 180 Å rods were subtracted from the respective distances in the other diameters. The s.d. of the distance shift distribution is a measure of the flexibility of the interaction. Boxes show SD with median line (line) and mean value (circle). Whiskers show the range within 1.5 IQR. Color code for residues: green=charged; yellow=hydrophobic; pink=polar+uncharged: grey=special cases. Color code for boxes: residues located in helix α1: red, residues located in helix α2 + 3: violet, residues located in helix α4: blue, residues located in helix α5: cyan. n = 11; n: Number of different rod diameters used for distance measurement. D: Diameters plotted against helical rise (after correction for number of strands) in Å. Apo (black): PspA ADP (red): PspA + 2 mM ADP. ATP (blue): PspA + 2 mM ATP. E: Diameters plotted against mass per 100 Å length in kDa. Apo (black): PspA. ADP (red): PspA + 2 mM ADP. ATP (blue): PspA + 2 mM ATP. F: PspA models with 60 monomers each show a decrease in rod length and an increasing diameter from 180 - 365 Å.
