ISDA: 12PB

ELECTRON MICROSCOPY

Sample

MTCH2 hyperactive mutant K25E Y235A V238D complexed with UCP1 nanobody

Specimen Preperation

Sample Aggregation State	PARTICLE
Vitrification Instrument	FEI VITROBOT MARK IV
Cryogen Name	ETHANE
Sample Vitrification Details	?

3D Reconstruction

Reconstruction Method	SINGLE PARTICLE
Number of Particles	612877
Reported Resolution (Å)	3.1
Resolution Method	FSC 0.143 CUT-OFF
Other Details	Final map generated by local refinement in cryoSPARC using a soft mask encompassing MTCH2 only.
Refinement Type
Symmetry Type	POINT

Map-Model Fitting and Refinement

ID	1
Refinement Space	REAL
Refinement Protocol	FLEXIBLE FIT
Refinement Target	Real-space correlation coefficient
Overall B Value	194
Fitting Procedure	?
Details	Initial fitting was done using Coot and refined by Phenix cryo-EM real space refinement.

Data Acquisition

Detector Type	GATAN K3 (6k x 4k)
Electron Dose (electrons/Å²)	60

Imaging Experiment

Date of Experiment	?
Temprature (Kelvin)
Microscope Model	TFS KRIOS
Minimum Defocus (nm)	800
Maximum Defocus (nm)	1800
Minimum Tilt Angle (degrees)	?
Maximum Tilt Angle (degrees)	?
Nominal CS	2.6
Imaging Mode	BRIGHT FIELD
Specimen Holder Model	FEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification	105000
Calibrated Magnification	?
Source	FIELD EMISSION GUN
Acceleration Voltage (kV)	300
Imaging Details	?

Imaging Experiment

Task	Software Package	Version
PARTICLE SELECTION	cryoSPARC	4.7.0
IMAGE ACQUISITION	SerialEM	?
CTF CORRECTION	cryoSPARC	4.7.0
MODEL FITTING	Coot	0.9.8.95
MODEL REFINEMENT	PHENIX	1.21.2_5419
INITIAL EULER ASSIGNMENT	cryoSPARC	4.7.0
FINAL EULER ASSIGNMENT	cryoSPARC	4.7.0
CLASSIFICATION	cryoSPARC	4.7.0
RECONSTRUCTION	cryoSPARC	4.7.0

Image Processing

CTF Correction Type	CTF Correction Details	Number of Particles Selected	Particle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTION	CTF parameters estimated using Patch CTF estimation in cryoSPARC. Global CTF correction for beam tilt was additionally performed.