Journal of Structural Biology, Vol.122, No.3, 267-282, 1998
Helical perturbations of the flagellar filament: Rhizobium lupini H13-3 at 13 angstrom resolution
Flagellar filaments are highly conserved structures in terms of the underlying symmetry of the polymer, subunit domain organization of the flagellin monomer, amino acid composition and primary sequence at the N and C termini. Traditionally, filaments are classified as "plain" or "complex." In complex filaments, the helical lattice is perturbed in a pairwise manner such that the symmetry is reduced along the 6-start helical lines. Both plain (unperturbed) and complex (helically perturbed) components are helically symmetric and share a common lattice. The perturbation in Rhizobium lupini H13-3 results in a subunit composed of a dimer of flagellin. We have generated a similar to 13 Angstrom resolution three-dimensional density map of the complex filament of R. lupini H13-3 from low-dose images of negatively stained filaments. Compared to a previous map, which extended to only similar to 25 Angstrom resolution and which was generated from only five filaments containing six layer-lines each, the current map is a product of merging 139 data sets containing 66 layer-lines each. The higher resolution and improved signal-to-noise yield a detailed and interpretable density map. The density map is divided into four concentric rings. These amount to two dense cylinders interconnected by low density radial spokes and wrapped by a three-start external winding. The unperturbed component of the map is strikingly similar to the known plain filament maps and, in particular, to that of Caulobacter crescentus. The helically perturbed component contributes mainly to the filaments's exterior (domain D3) where it comprises the tips of the outer domains interconnecting, pairwise, along the 11-start protofilaments and, again, laterally along the 6-start lines forming vertical and horizontal loops. Strong intersubunit connectivity occurs in the D2 shell and in the inner shell which is dominated by 3-start densities. The contribution of the complex component to the radial spokes seems negligible.
Keywords:complex bacterial flagellar filament;electron microscopy;helical perturbations;Rhizobium lupini H13-3;three-dimensional helical reconstruction