6W98

Single-Particle Cryo-EM Structure of Arabinofuranosyltransferase AftD from Mycobacteria


ELECTRON MICROSCOPY
Sample
Wild-type Mycobacterial Arabinofuranosyltransferase AftD Complexed with Acyl Carrier Protein
Sample Components
F5/8 type C domain-containing protein
Acyl carrier protein
Specimen Preparation
Sample Aggregation StatePARTICLE
Vitrification InstrumentSPOTITON
Cryogen NameETHANE
Sample Vitrification DetailsThe Spotiton V1.0 robot was operating at room temperature and moderate humidity.
3D Reconstruction
Reconstruction MethodSINGLE PARTICLE
Number of Particles490616
Reported Resolution (Å)2.9
Resolution MethodFSC 0.143 CUT-OFF
Other Details
Refinement Type
Symmetry TypePOINT
Point SymmetryC1
Map-Model Fitting and Refinement
Id1
Refinement SpaceREAL
Refinement ProtocolAB INITIO MODEL
Refinement Target
Overall B Value
Fitting Procedure
Details
Data Acquisition
Detector TypeGATAN K2 QUANTUM (4k x 4k)
Electron Dose (electrons/Å**2)95.72
Imaging Experiment1
Date of Experiment
Temperature (Kelvin)
Microscope ModelFEI TITAN KRIOS
Minimum Defocus (nm)1000
Maximum Defocus (nm)2000
Minimum Tilt Angle (degrees)
Maximum Tilt Angle (degrees)
Nominal CS2.7
Imaging ModeBRIGHT FIELD
Specimen Holder ModelFEI TITAN KRIOS AUTOGRID HOLDER
Nominal Magnification130000
Calibrated Magnification
SourceFIELD EMISSION GUN
Acceleration Voltage (kV)300
Imaging Details
EM Software
TaskSoftware PackageVersion
PARTICLE SELECTIONGautomatch0.53
IMAGE ACQUISITIONLeginon3.4
CTF CORRECTIONRELION2.1
CTF CORRECTIONcisTEM1
CTF CORRECTIONcryoSPARC2
MODEL FITTINGCoot
MODEL FITTINGBuccaneer
MODEL REFINEMENTPHENIX
INITIAL EULER ASSIGNMENTcryoSPARC2
FINAL EULER ASSIGNMENTcryoSPARC2
CLASSIFICATIONRELION2.1
RECONSTRUCTIONRELION2.1
Image Processing
CTF Correction TypeCTF Correction DetailsNumber of Particles SelectedParticle Selection Details
PHASE FLIPPING AND AMPLITUDE CORRECTIONCTF was estimated using GCTF and refined throughout the pipeline using cisTEM.2800102