5A79

Novel inter-subunit contacts in Barley Stripe Mosaic Virus revealed by cryo-EM

ELECTRON MICROSCOPY

Sample
CHIMERIC BARLEY STRIPE MOSAIC VIRUS WIDE VIRION

Specimen Preparation
Sample Aggregation State	FILAMENT
Vitrification Instrument	HOMEMADE PLUNGER
Cryogen Name	ETHANE
Sample Vitrification Details	VITRIFICATION 1 -- CRYOGEN- ETHANE, HUMIDITY- 40, TEMPERATURE- 100, INSTRUMENT- HOMEMADE PLUNGER, METHOD- 3.5UL OF SAMPLE WAS ADDED TO CONTINUOUS CARB ...	VITRIFICATION 1 -- CRYOGEN- ETHANE, HUMIDITY- 40, TEMPERATURE- 100, INSTRUMENT- HOMEMADE PLUNGER, METHOD- 3.5UL OF SAMPLE WAS ADDED TO CONTINUOUS CARBON COATED OR C-FLAT GRIDS, BLOTTED FOR 2 SECONDS AND THEN PLUNGED IN LIQUID ETHANE,

3D Reconstruction
Reconstruction Method	SINGLE PARTICLE
Number of Particles	2077
Reported Resolution (Å)	4.1
Resolution Method
Other Details	DISORDERED REGIONS WERE MODELED STEREOCHEMICALLY
Refinement Type
Symmetry Type	HELICAL

Map-Model Fitting and Refinement
Id	1
Refinement Space	REAL
Refinement Protocol	FLEXIBLE FIT
Refinement Target	Cross-correlation coefficient
Overall B Value
Fitting Procedure
Details	METHOD--RIGID BODY USING CHIMERA, FOLLOWED BY FLEXIBLE FITTING IN FLEX-EM AND COOT. THEN REALSPACE REFINEMENT IN PHENIX FOLLOWED BY MANUAL ADJUSTMENT ...	METHOD--RIGID BODY USING CHIMERA, FOLLOWED BY FLEXIBLE FITTING IN FLEX-EM AND COOT. THEN REALSPACE REFINEMENT IN PHENIX FOLLOWED BY MANUAL ADJUSTMENT IN COOT (INTERATIVE) REFINEMENT PROTOCOL--CRYO-EM

Data Acquisition
Detector Type	GATAN ULTRASCAN 4000 (4k x 4k)
Electron Dose (electrons/Å**2)	25

Imaging Experiment	1
Date of Experiment	2012-06-01
Temperature (Kelvin)	90
Microscope Model	FEI TECNAI F30
Minimum Defocus (nm)	700
Maximum Defocus (nm)	3500
Minimum Tilt Angle (degrees)
Maximum Tilt Angle (degrees)
Nominal CS	2.3
Imaging Mode	BRIGHT FIELD
Specimen Holder Model
Nominal Magnification	150000
Calibrated Magnification	150000
Source	FIELD EMISSION GUN
Acceleration Voltage (kV)	300
Imaging Details

EM Software
Task	Software Package	Version
RECONSTRUCTION	EMAN
RECONSTRUCTION	IMAGIC
RECONSTRUCTION	SPIDER

Image Processing
CTF Correction Type	CTF Correction Details	Number of Particles Selected	Particle Selection Details
	PHASE FLIPPED EACH PARTICLE

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.