1ML5

Structure of the E. coli ribosomal termination complex with release factor 2

ELECTRON MICROSCOPY

Sample
release factor RF2 bound to E. coli ribosomes
Sample Components
peptide release factor RF2

Specimen Preparation
Sample Aggregation State	PARTICLE
Vitrification Instrument	HOMEMADE PLUNGER
Cryogen Name	ETHANE
Sample Vitrification Details	holey carbon grid at 20C; flash frozen into liquid ethan

3D Reconstruction
Reconstruction Method	SINGLE PARTICLE
Number of Particles	15800
Reported Resolution (Å)	14
Resolution Method
Other Details	iterative refinement procedure as described in Quat. Rev. of Biophysics 33, 4 (2000), pp. 307-369
Refinement Type
Symmetry Type	POINT
Point Symmetry	C1

Map-Model Fitting and Refinement
Id	1 (1GQE, 1GIX, 1GIY)
Refinement Space	REAL
Refinement Protocol	RIGID BODY FIT
Refinement Target	best visual fit using the program O
Overall B Value
Fitting Procedure
Details	REFINEMENT PROTOCOL--rigid body fit of the 70S ribosome without individual fitting of proteins or RNA fragments. Rigid body fit of three individual ...	REFINEMENT PROTOCOL--rigid body fit of the 70S ribosome without individual fitting of proteins or RNA fragments. Rigid body fit of three individual domains of release factor RF2 keeping the connectivity; linkers betweenn domains defined based on temperature factors of crystal structure, conserved Gly and Pro residues, proteolytic sites, and global domain architecture

Data Acquisition
Detector Type	KODAK SO-163 FILM
Electron Dose (electrons/Å**2)	10

Imaging Experiment	1
Date of Experiment	2001-09-06
Temperature (Kelvin)	100
Microscope Model	FEI/PHILIPS CM200FEG/ST
Minimum Defocus (nm)	500
Maximum Defocus (nm)	2900
Minimum Tilt Angle (degrees)
Maximum Tilt Angle (degrees)
Nominal CS	2.1
Imaging Mode	BRIGHT FIELD
Specimen Holder Model
Nominal Magnification	50000
Calibrated Magnification	48000
Source	FIELD EMISSION GUN
Acceleration Voltage (kV)	200
Imaging Details	low dose mode

Image Processing
CTF Correction Type	CTF Correction Details	Number of Particles Selected	Particle Selection Details
	phase flipping CTF correction of each particle as function of position in the micrograph

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.