L-type straight flagellar filament made of full-length flagellin
ELECTRON MICROSCOPY
Sample
L-TYPE STRAIGHT FLAGELLAR FILAMENT
Specimen Preparation
Sample Aggregation State
FILAMENT
Vitrification Instrument
Cryogen Name
ETHANE
Sample Vitrification Details
3D Reconstruction
Reconstruction Method
HELICAL
Number of Particles
Reported Resolution (Å)
4
Resolution Method
Other Details
The layer-line spacing of the L-type straight flagellar filament is more than twice longer than that of the R-type, and frozen-hydrated L-type filamen ...
The layer-line spacing of the L-type straight flagellar filament is more than twice longer than that of the R-type, and frozen-hydrated L-type filaments tended to be more curved. Hence, straightening of images using bi-cubic spline was carried out at the first stage. The three-dimensional map was calculated by helical image reconstruction. An initial map produced from 70 filament images, corresponding to 51,272 molecular images, showed relatively poor connections of the ND0 helix and the NS loop. Hence, images showing an inter-particle phase residual against the initial average larger than 40 were excluded from the data set to be averaged. As a result, a three-dimensional map of a better quality was obtained. The number of filament images used for the final image reconstruction was 47, and the total number of molecular images was 36,842, which is consistent with an estimated number of molecular images required to achieve this resolution range.
Refinement Type
Symmetry Type
HELICAL
Map-Model Fitting and Refinement
Id
1
Refinement Space
RECIPROCAL
Refinement Protocol
FLEXIBLE FIT
Refinement Target
AMPLITUDE-WEIGHTED PHASE RESIDUAL
Overall B Value
Fitting Procedure
Details
REFINEMENT PROTOCOL--POSITIONAL AND SIMULATED ANNEALING
