ISDA: 13HE

ELECTRON MICROSCOPY

Sample

AdhE spirosome

Specimen Preperation

Sample Aggregation State	FILAMENT
Vitrification Instrument	FEI VITROBOT MARK IV
Cryogen Name	ETHANE
Sample Vitrification Details	Vitification was performed on CFlat 1.2/1.3 300 mesh holey carbon grids with copper support.

3D Reconstruction

Reconstruction Method	HELICAL
Number of Particles	95407
Reported Resolution (Å)	4.07
Resolution Method	FSC 0.143 CUT-OFF
Other Details	Overall resolution 4.07 angstrom as estimated by the masked gold-standard FSC = 0.143 criterion in cryoSPARC. Unmasked FSC values are lower, consistent with expected effects of masking." in the details field.
Refinement Type
Symmetry Type	HELICAL

Map-Model Fitting and Refinement

ID	1
Refinement Space	?
Refinement Protocol	RIGID BODY FIT
Refinement Target	?
Overall B Value	?
Fitting Procedure	?
Details	A existing structure's (PDB:8UHW) coordinates were relaxed using the Rosetta Fast Relax algorithm and refined against the EM density from cryoSPARC using Phenix and Coot. Histidine residues coordinate the iron ion; short distances flagged as clashes correspond to metal coordination geometry.

Data Acquisition

Detector Type	GATAN K3 (6k x 4k)
Electron Dose (electrons/Å²)	48

Imaging Experiment

Date of Experiment	?
Temprature (Kelvin)
Microscope Model	TFS KRIOS
Minimum Defocus (nm)	100
Maximum Defocus (nm)	500
Minimum Tilt Angle (degrees)	?
Maximum Tilt Angle (degrees)	?
Nominal CS	?
Imaging Mode	BRIGHT FIELD
Specimen Holder Model	?
Nominal Magnification	130000
Calibrated Magnification	?
Source	FIELD EMISSION GUN
Acceleration Voltage (kV)	300
Imaging Details	?

Imaging Experiment

Task	Software Package	Version
PARTICLE SELECTION	cryoSPARC	4.4.1
CTF CORRECTION	cryoSPARC	4.4.1
MODEL FITTING	Coot	0.9.8.96
INITIAL EULER ASSIGNMENT	cryoSPARC	4.4.1
FINAL EULER ASSIGNMENT	cryoSPARC	4.4.1
RECONSTRUCTION	cryoSPARC	4.4.1
MODEL REFINEMENT	PHENIX	2.0-5936

Image Processing

CTF Correction Type	CTF Correction Details	Number of Particles Selected	Particle Selection Details
NONE	?