5J4D

E. coli release factor 1 bound to the 70S ribosome in response to a pseudouridylated stop codon

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL12-2
Synchrotron site	SSRL
Beamline	BL12-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2015-07-26
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9795
Spacegroup name	P 21 21 21
Unit cell lengths	211.541, 454.397, 619.473
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.977 - 3.100
R-factor	0.2221
Rwork	0.221
R-free	0.25630
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	1.072
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall
Low resolution limit [Å]	70.000
High resolution limit [Å]	3.100
Number of reflections	1174254
<I/σ(I)>	8.28
Completeness [%]	100.0
Redundancy	37.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	3.1 uL 70S*mRNA*tRNAfMet complex + 3.1 uL crystallization buffer (0.1 M Tris-HCl, pH 7.5, 4% v/v PEG20000, 8% v/v MPD, 0.2 M potassium thiocyanate), reservoir: 300 uL 0.5-0.7 M sodium chloride, 4-fold molar excess of E. coli release factor 1 added during complex formation
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	3.1 uL 70S*mRNA*tRNAfMet complex + 3.1 uL crystallization buffer (0.1 M Tris-HCl, pH 7.5, 4% v/v PEG20000, 8% v/v MPD, 0.2 M potassium thiocyanate), reservoir: 300 uL 0.5-0.7 M sodium chloride, 4-fold molar excess of E. coli release factor 1 added during complex formation
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	3.1 uL 70S*mRNA*tRNAfMet complex + 3.1 uL crystallization buffer (0.1 M Tris-HCl, pH 7.5, 4% v/v PEG20000, 8% v/v MPD, 0.2 M potassium thiocyanate), reservoir: 300 uL 0.5-0.7 M sodium chloride, 4-fold molar excess of E. coli release factor 1 added during complex formation
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	3.1 uL 70S*mRNA*tRNAfMet complex + 3.1 uL crystallization buffer (0.1 M Tris-HCl, pH 7.5, 4% v/v PEG20000, 8% v/v MPD, 0.2 M potassium thiocyanate), reservoir: 300 uL 0.5-0.7 M sodium chloride, 4-fold molar excess of E. coli release factor 1 added during complex formation
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	3.1 uL 70S*mRNA*tRNAfMet complex + 3.1 uL crystallization buffer (0.1 M Tris-HCl, pH 7.5, 4% v/v PEG20000, 8% v/v MPD, 0.2 M potassium thiocyanate), reservoir: 300 uL 0.5-0.7 M sodium chloride, 4-fold molar excess of E. coli release factor 1 added during complex formation
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	3.1 uL 70S*mRNA*tRNAfMet complex + 3.1 uL crystallization buffer (0.1 M Tris-HCl, pH 7.5, 4% v/v PEG20000, 8% v/v MPD, 0.2 M potassium thiocyanate), reservoir: 300 uL 0.5-0.7 M sodium chloride, 4-fold molar excess of E. coli release factor 1 added during complex formation
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	3.1 uL 70S*mRNA*tRNAfMet complex + 3.1 uL crystallization buffer (0.1 M Tris-HCl, pH 7.5, 4% v/v PEG20000, 8% v/v MPD, 0.2 M potassium thiocyanate), reservoir: 300 uL 0.5-0.7 M sodium chloride, 4-fold molar excess of E. coli release factor 1 added during complex formation