ISDA: 9R15

ELECTRON MICROSCOPY

Sample

Bovine complex I at pH 9.2 in the deactive state (consensus)

Specimen Preperation

Sample Aggregation State	PARTICLE
Vitrification Instrument	FEI VITROBOT MARK IV
Cryogen Name	ETHANE
Sample Vitrification Details	Blotted with a force of -10 for 10 seconds prior to plunge freezing.

3D Reconstruction

Reconstruction Method	SINGLE PARTICLE
Number of Particles	262659
Reported Resolution (Å)	2.31
Resolution Method	FSC 0.143 CUT-OFF
Other Details	?
Refinement Type
Symmetry Type	POINT

Map-Model Fitting and Refinement

ID	1
Refinement Space	REAL
Refinement Protocol	RIGID BODY FIT
Refinement Target	Cross-correlation coefficient
Overall B Value	?
Fitting Procedure	?
Details	Initial rigid-body fitting was done using UCSF ChimeraX fit in map tool, with modifications made in Coot, followed by successive rounds of real-space refinement in PHENIX.

Data Acquisition

Detector Type	GATAN K3 BIOQUANTUM (6k x 4k)
Electron Dose (electrons/Å²)	40.50

Imaging Experiment

Date of Experiment	?
Temprature (Kelvin)
Microscope Model	TFS KRIOS
Minimum Defocus (nm)	900
Maximum Defocus (nm)	2300
Minimum Tilt Angle (degrees)	?
Maximum Tilt Angle (degrees)	?
Nominal CS	2.7
Imaging Mode	BRIGHT FIELD
Specimen Holder Model	FEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification	81000
Calibrated Magnification	?
Source	FIELD EMISSION GUN
Acceleration Voltage (kV)	300
Imaging Details	?

Imaging Experiment

Task	Software Package	Version
PARTICLE SELECTION	RELION	4.0-BETA
IMAGE ACQUISITION	EPU	?
CTF CORRECTION	RELION	4.0-BETA
CTF CORRECTION	cryoSPARC	3.3.2
MODEL FITTING	UCSF ChimeraX	1.7
MODEL FITTING	Coot	0.9.8.1
INITIAL EULER ASSIGNMENT	RELION	4.0-BETA
FINAL EULER ASSIGNMENT	cryoSPARC	3.3.2
CLASSIFICATION	cryoSPARC	3.3.2
RECONSTRUCTION	cryoSPARC	3.3.2
MODEL REFINEMENT	PHENIX	1.21_5207

Image Processing

CTF Correction Type	CTF Correction Details	Number of Particles Selected	Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION	Global and local CTF parameters were refined both in cycles in RELION then during refinement cycles in the final homogeneous refinement in cryoSPARC.